CA2196806A1 - Polynucleotide reagents having nonnucleotidic moieties, and associated methods of synthesis and use - Google Patents
Polynucleotide reagents having nonnucleotidic moieties, and associated methods of synthesis and useInfo
- Publication number
- CA2196806A1 CA2196806A1 CA002196806A CA2196806A CA2196806A1 CA 2196806 A1 CA2196806 A1 CA 2196806A1 CA 002196806 A CA002196806 A CA 002196806A CA 2196806 A CA2196806 A CA 2196806A CA 2196806 A1 CA2196806 A1 CA 2196806A1
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- Canada
- Prior art keywords
- group
- alkyl
- reagent
- hydrogen
- dna
- Prior art date
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- Abandoned
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
- C07H15/02—Acyclic radicals, not substituted by cyclic structures
- C07H15/04—Acyclic radicals, not substituted by cyclic structures attached to an oxygen atom of the saccharide radical
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
- C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic System
- C07F9/02—Phosphorus compounds
- C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
- C07F9/655—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having oxygen atoms, with or without sulfur, selenium, or tellurium atoms, as the only ring hetero atoms
- C07F9/65515—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having oxygen atoms, with or without sulfur, selenium, or tellurium atoms, as the only ring hetero atoms the oxygen atom being part of a five-membered ring
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6813—Hybridisation assays
- C12Q1/6816—Hybridisation assays characterised by the detection means
- C12Q1/6823—Release of bound markers
Abstract
Methods and reagents are provided for synthesizing polynucleotides containing modified deoxyribose residues. Monomeric reagents having structural formula (I), wherein R1, R2, R3, R4 and R5 are as defined herein, are used to create polynucleotides having nonnucleotidic moieties -A-Z-(R9)n at the 1 position of selected deoxyribose units. The polynucleotides so provided are useful in a variety of hybridization assay formats.
Description
2 1 ~
Polynucleotide Reagents having nonnucleotidic moieties, and associated methods of synthesis and use.
5 Tf rhnir:~l Field This invention relates generally to nucleic acid chemistry, i.e., DNA synthesis,hyl"iJ;~Liul~ assays, and the like, and to reagents used in conjunction therewith. More partiCularly~ the invention relates to methods and monomeric reagents for rntroducing sites--containing modified d~ ;I,u~ .O;.,..~, pu~ cu~ . The rnvention additionally relates to methods of using the monomeric reagents of the invention and pcl~ ,R,uL;dc reagents synthesized therefrom in DNA hyl)lil;~liull assays.
Backvrrl~
Nucleic acid l~Jb.i.li ~iO~I assays are commonly used in genetic research, biomedical research and clinical diagnostics. In a basic nucleic acid hJI).i.li~Liull assay, the nucleic acid of interest is hybridized, in single-stranded forrn, to a labeled srngle-stranded nucleic acid probe and resultmg labeled duplexes are detected. Variations of this basic scheme have been developed to enhance accuracy, facilitate the separation of the duplexes to be detected from extraneous materials, and/or amplify the signal that is detected.
2 0 Commonly assignGd U. S . Patent No. 5,43û, 13 6, ill~,UI ~IUI _a,1 by reference herein, describes a tectmique whereby selectably cleavable sites are introduced into ~,':,,...,... 1, ..I;Sf chains, enabling release of a detectable label after hJ.JIill;~Liull is complete. As explained in that application, selectably cleavable sites are useful in a number of different types of hJbli~ iUII assay formats. For example, in one type of assay rn which hJbli~ Livll gives 25 rise to a ' ' s~.l",u. ~cl duplex of a labeled probe and sample DNA, a selectably cleavable site contained within the hybrid structure will enable ready separation of the label from the solid support. Commonly assigned U.S. Patent Nos. 4,775,619 and 5,118,605 are respectively directed to the use of restriction _...1. ,... Irl__~_ cleavable sites in such assays and the use of chemically cleavable sites (e.g., disulfide linkages, 1,2-diols, and the like). These 3 0 cleavable sites cam be introduced during . '~ 1 ,1; 1r- synthesis, and are cleavable with restriction ,"rL ,. ,... 1. A - . in the case of restriction sites and with particular chemical reagents, e.g., with thiols, periodate, or the like, in the case of chemically cleavable sites.
Polynucleotide Reagents having nonnucleotidic moieties, and associated methods of synthesis and use.
5 Tf rhnir:~l Field This invention relates generally to nucleic acid chemistry, i.e., DNA synthesis,hyl"iJ;~Liul~ assays, and the like, and to reagents used in conjunction therewith. More partiCularly~ the invention relates to methods and monomeric reagents for rntroducing sites--containing modified d~ ;I,u~ .O;.,..~, pu~ cu~ . The rnvention additionally relates to methods of using the monomeric reagents of the invention and pcl~ ,R,uL;dc reagents synthesized therefrom in DNA hyl)lil;~liull assays.
Backvrrl~
Nucleic acid l~Jb.i.li ~iO~I assays are commonly used in genetic research, biomedical research and clinical diagnostics. In a basic nucleic acid hJI).i.li~Liull assay, the nucleic acid of interest is hybridized, in single-stranded forrn, to a labeled srngle-stranded nucleic acid probe and resultmg labeled duplexes are detected. Variations of this basic scheme have been developed to enhance accuracy, facilitate the separation of the duplexes to be detected from extraneous materials, and/or amplify the signal that is detected.
2 0 Commonly assignGd U. S . Patent No. 5,43û, 13 6, ill~,UI ~IUI _a,1 by reference herein, describes a tectmique whereby selectably cleavable sites are introduced into ~,':,,...,... 1, ..I;Sf chains, enabling release of a detectable label after hJ.JIill;~Liull is complete. As explained in that application, selectably cleavable sites are useful in a number of different types of hJbli~ iUII assay formats. For example, in one type of assay rn which hJbli~ Livll gives 25 rise to a ' ' s~.l",u. ~cl duplex of a labeled probe and sample DNA, a selectably cleavable site contained within the hybrid structure will enable ready separation of the label from the solid support. Commonly assigned U.S. Patent Nos. 4,775,619 and 5,118,605 are respectively directed to the use of restriction _...1. ,... Irl__~_ cleavable sites in such assays and the use of chemically cleavable sites (e.g., disulfide linkages, 1,2-diols, and the like). These 3 0 cleavable sites cam be introduced during . '~ 1 ,1; 1r- synthesis, and are cleavable with restriction ,"rL ,. ,... 1. A - . in the case of restriction sites and with particular chemical reagents, e.g., with thiols, periodate, or the like, in the case of chemically cleavable sites.
4 ~ 6~6 -2~ I //o The present invention is also directed in part to the in~,ul~uul~Liull of selectably cleavable sites into ~ul~..ucl~,v~;~L,.,. The cleavable sites herein are contained within a linker arm present at the I position of a Jcv~yl iL,ùse molecule. In addition to providing such cleavable sites. the invention also relates to the creation of "abasic sites" within 5 pvl~u~ id~", i.e.. monomeric units which contain the d.,~J~.yli1Oa~. ring but do not have a purine or pyrimidine base present at the I position. Such abasic sites are useful in a wide variety of contexts, as will be explained in detail 1.... c;..' ch, ... For example, an abasic site may be used to create branched DNA, i.e., a multimeric poly... cl~,vtilL, structure in which three poly..u~.lcv~iJc chains emanate from a single uev~.ylibuac unit. These branch points are 1 0 extremely useful in providing large, "---ul~i_.i-," DNA structures which cam then be used im assays. Abasic sites may also be used in other ways, e.g., in the synthesis of DNA bound to a solid support (typically although not necessarily at the I position), to reverse the direction of chemical DNA synthesis, i.e., 3'~5' to 5' ~3' or vice versa, and in triple heLx formation.
Thus, in addition to utility in providing cleavable sites wjthin, 'i~ ' ' or l)ul~..u~ ,v~ide chains, the invention enables a number of procedures deriving from the presence of linker arms at the I position of a monomeric dcv~yl ib~/a~. unit rather than purine or pyrimidine bases as present in cu~ liu~ nucleotide structures.
2 0 Overview of thP Art Backgrûund references which relate generally to methods for bylllh~;~
.,1;5 """ l~vl~' include those related to 5'-to-3' syntheses based on the use of,B-cyanoethyl phosphate protecting groups, e.g., de Napoli et al., Gazz Chim rt~l 114:65 (1984), Rosenthal et aL, Tetrrhp~rûn T .PttPrS 24: 1691 (1983),1Belagaje and Brush, Nucleic ~ri~lc F PCP~rrh 10:6295 (1977), in references which describe solution-phase 5'-to-3' syntheses include Hayatsu and Khorana, J American ~'hPmir~l Society 89:3880 (1957), Gait amd Sheppard, ~llrlPir Aririe RP~P~rrh 4 1135 (1977), Cramer and Koster, Aneew ('hPm Int E.l Fn~d _:473 (1968), and Blackbum et al., Journal ofthe Chemical Society, Part C, 2438 (1967).
In addition to the above-cited art, Matteucci and Caruthers, J. AmPrir~n ~hP.n;. ~1 Society 103:3185-3191 (1981), describe the use of ~ .h. ~ in the preparation of r~ 1 vl; !~ ~ Beaucage and Caruthers, Tetrahedron Letters;~:1859-1862 (1981), and U S Patent No 4~415~732 describe the use of l l ~ in the preparation of . _, .. , ., .. , .. . _ , _ , , 2~96806 '~ r s ~ WO96/06104 3 PCT/US95/10776 . vL; !r~ Smith, ABL 15-24 (December 1983), describes automated solid-phase o:;gvJ~v~yl il ;-, ,. .1, . .~h;P synthesis. See also the references cited therein, and Warner et al., DNA 3 :401411 (1984), whose disclosure is h~,u~5~u~ J herein by reference.
U.S. Patent Nos. 4,483,964 and 4,517,338 to Urdea et al. describes a method for S ~y..~h.,~ g pol~"...,l~,vfid~,s by selectively introducing reagents to a solid phase substrate in a tubular reaction zone. U.S. Patent No. 4,910,300 to Horn et al. also describes a method for ~y~lih.,~ g rl;r","." If . ,I;s~ - by sequentially adding nucleotidic monomers to a growing chain, but involves the in~ùl ~u~ ~L;ull of labelled, N-4 modified cytosine residues at ~-~d~ ... J, spaced apart positions. U.S. Patent No. 5,256,549 to Horn et al. is also of 1 û interest in that a method for preparing .~ f vl;: - is provided which involves a -6. .,. technique, i.e., in which the desired ,~ . ,"",.1. v~ is essentially synthesized and "purified" ~ f. ~ ly~ such that the final product is produced in ~ , pure form.
HomandUrdea,DNA5(5):421-425(1986),describe~uhv~vhvlyl~Liullofsolid-15 supported DNA fragments using bis(~ ..u~il.w~y)-N,N-diisopropyl ,t ~ See also, Horn and Urdea, Tetrahedron T PtfPr~ 27:47054708 (1986).
References which relate to hybl;J;~L;ùn techniques in general include the following:
Meinkoth and Wahl, Anal. Biol '...,. ny 138:267-284 (1984), provide an excellent review of hys,l;J;~L;u~ techniques. Leary et al., Proc. Natl. Ar Irl Sci. ~fUSA) 80:4045-4049 (1983) 2 0 describe the use of biotinylated DNA in conjunction with an avidin-enzyme conjugate for detectionofspecifico!;,~v,,lcl~.vL;Jesequences. R~nkietal.,Gene21:77-85,describewhat they refer to as a "sandwich" hylJI;J;~L;ull for detection of, .I;g. ,~ sequences.
Pfeuffer and Helmrich, J. Biol. Chem. ~:867-876 (1975), describe the coupling ofguanosine-5'-0-(3-i'i . ' - . ' ) to Sepharose 4B. Bauman et al., J. Hi~fr r hPm ~n~
Cvtochem 29:227-237, describe the 3'-labeling of RNA with fluorescers. PCT Application 7 describes the addition to DNA fragments of modified l;b.. If vl ;. l. for labeGng and methods for analyzing such DNA fragments. Renz and Kurz, Nucl. Ari~c Rr~
12:3435-3444, describe the covalent linking of enzymes to u~ ; ' - Wallace, DNA
R~ ~ T~ Woo, S., ed.) CRC Press, Boca Raton, Florida, provides a 3 0 general background of the use of probes in diagnosis. Chou and Merigan, N. ~;n,p J of ~L 308 921-925, describe the use of a ~ JI~5~e-labeled probe for the detection of CMV. Lnm3r~ M,-fhr,l~ in Fn7vmol 34B, 24:77-102 (1974), describes procedures for Wos6/06io4 &~S6 ~ //6 linking to poly~,.,.yhu~l;d~,." while Parikh et al., Methods in Enzymol. 34B, 24:77-102 (1974) describe coupling reactions with agarose. Aiwine et al., Proc. Natl. Acad. Sci. (USA) 74:5350-5354 (1977), describe a method of transferring .~ from gels to a solid support for hyblici;LGl;ull. Chu et al., Proc. Natl. Acad. Sci. (USA) 11 :6513-6529, describe a technique for derivatizing temlinal nucleotides. Ho et ai., F~ . .h. . . ,;~l ~y 20:64-67 (1981), describe derivatizing temlinal nucleotides through phosphate to fomm esters. Ashiey and MacDonaid, Anai. Bionh~m 140.95-103 (1984), report a method for preparing probes from a surface-bound template.
Home and Dervan, J. Am. Chem. Soc I i2:2435-2437 (1990), and Froehier et ai., Bi(lrh~mi~hv 31: 1603- 1 ~)9 (1992), relate to . .1;.~ r vl ;- Ie-directed triple helix formation.
Summarv of the Inv~ nti~ln in one aspect of the invention, then7 monomeric reagents useful for providing the 15 novel polyllu~levL;de structures are provided, the monomeric reagents having the structural fommula (I) ~ R4 ~ ~Rs wherein:
R' is selected from the group consisting of hydrogen, acid-sensitive, base-stable protecting groups and acyl capping groups;
R2 is a phosphorus derivative selected to enable addition of the reagent to a 30 molecular species containing a free hydroxyl group, or is a linkage to a solid support;
R3 is selected from the group consisting of hydrogen, hydroxyl, sulfhydryl, halogeno, amino, aikyl, allyl~ oR6 wherein R6 is alkyl, allyl, silyl or phosphate;
~ WO96/06104 19680~ s ' r~ 6 R4 is either hydro~en or -(CH2)mOR7 wherein R' is alicyl or -(Co)R5, R5 is alL-yl, and m is an integer in the range of 0 to 12 inclusive;
Rs is ~A~Z~X(R9)n;
A is oxygen, sulfur or methylene;
~ 5 Z is arylene, C6-C,5 aralicylene or C~-C~z alkylene containing 0 to 6 I.~,Ltlu~.Lu,.. .
selected from the group consisting of O, S, N, Si and Se and 0 to 6 linL-ages selected from the group consisting of -CO-,-COO-, -CONH-, -NHCO-, -S-S-, -SOz-, -CH(OH)-CH(OH)-, -CH(oR4)-CH(oR4)-, -O-PO(O~-O-, -o-Po(R4)-,-o-Po(oR4)-o-7 -o-Po(oR4)-R5- and -Po(oR4)-o-R5- in which R4 is lower aiLyl and R5 is lower alLylene, and, if Z is aralLylene or 10 allylene, containing 0 to 3 unsaturated bonds;
X is selected from the group consisting of-NH-, -CONH-, -NHCO-, -CO-, -S- and -SiE;
R9 is hydrogen, a protecting group, a detectable label, or, uniess X is _SjE, a solid support; and n is I when X is -NH-, -CONH-, -NHCO-, -CO-, or -S-, and is 3 when X is -Si=.
In another aspect, ~oir..,~ ,Li i~ reagents are provided having the structurai formulae (II), (III) or (IV) ~ ~lo ~ o--P--" ~ R
Rs . O R~
2 5 11 o--P--o-- ~ ~ to A 1--~
a r ~9~Q6 WO 96106104 - 6 - - - r~ O
s ~C--p_ ~ 4 (111) O~ R
5'- ~ c-&~o~ 4 ~1~ \~p~
(IV) ~ P-O -~ IDN~ ~S-C~
wherein DNA, represents a first segment of DNA DNA~ represents a second segment of DNA and R3, R~ and R5 are as defined above. In a related aspect of the invention, branched DNA is provided having the structural formula (V) WO96/06104 9680~ 7 r~l~u..,J,i l16 1~~ ~hlA~ ~- c - ~- O ~
~ CA ~ a l'l A ~ 1-- ~ l~
~ ~ lP o ~ o ~.~
10 wherein DNA,, DNA2 and DNA3 represent firso second and third segments of DNA, and R;, R', A and Z are as defined above.
In still other aspects of the invention, methods are provided for synthesizing pcl~ iJ~ containing abasic sites and for preparing branched DNA. These methods involve the ;-,~u- ~,u, 4L;UII of the above-mentioned monomeric reagent into larger 15 p~ ..u~,L,uL;de structures.
A method is also provided for detecting the presence of an ~I g- ~ u~ e sequenceof interest in a sample which involves hybridizing the nucleic acid sample with a pGlJ"~,leolide probe containing an abasic site as described herein, wherein the abasic site is formed from the monomeric reagent defined above, and fiurther wherein the reagent contains 2 0 a detectable label at R3 and a cleavable site within the linker moiety -Z-. Either the sample or the po .~ ,vLide probe is bound to a solid support, such that hybridization results in a label being bound to the support through the cleavable site. Following l,yb, ;.1;~4fio-" the cleavable site is cleaved with a suitable reagent so as to release the detectable label R3, and label which is free of the support is quantitated and correlated with the presence andlor quantity of 2 5 sample.
Additionally, probes synthesized using the compounds of the invention may contain 3'-3' linkages, as illustrated in structures (Ill) and (IV) above. Ol;6oue~J~.yllu~l~ul;ll~ probes containing 3'-3' linkages can be used in triple helix formation, i.e., as such probes can bind to opposite strands of duplex DNA.
WO96/06104 2~ 96~~6 - 8i - P~ /6 1 pPt~ i Descrivtion of the Invention Definitions and .~. ," .. ,1 l ". ,:
Before the present invention is disclosed and described in detail, it is to be understood that this invention is not iimited to specific assay formats, materials or reagents, 5 as such may, of course, vary. It is aiso to be understood that the i ~ vJ used herein is for the purpose of describing particular r ~1 ~0/~ oniy and is not intended to be limiting.
It must be noted that, as used in the ~ ;..,. amd the appended claims, the singular forms "a," "an" and "the" include plurai referents uniess the context clearly dictates otherwise. Thus, for example, reference to "a monomeric reagent" includes mixtures of 10 monomeric reagents, reference to "a pu4..l~,1evL;de probe" may include mixtures of different probes, reference to a po4..~1vlLvLiJc containing "an abasic site" includes l~u4~u~,lvvLi;lC~
containing two or more abasic sites, and the like.
In this ~ .", and in the claims which follow, reference will be made to a number of terms which shail be defined to have the following meanings:
As used herein, the terms "~u4.~ evLi ie'~ and ". 1~ " shail be generic to pcl~J~vAyl ;1~ L ..1;.5. ~ (containing 2-deoxy-D-ribose), to ~uly. ;1..., . 1 .1; 1. (containing D-ribose), to any other type of po4..v~ Iev~ide which is an N-giycoside of a purine or pyrimid;ne base, and to other polymers containing ~ backbones (e.g., protein nucleic acids and synthetic sequence-specific nucleic acid polymers vUIIIII~ available 2 0 from the Anti-Gene Development Group, Corvailis, Oregon, as NeugeneTM polymers), providing that the polymers contain .,. ,. 1~ ob ~-~ in a ~ 5v ~iun which ailows for base pairing and base stacking, such as is found in DNA and RNA. There is no intendeddistinction in length between the term "I,c~l~.lJ~ .vLi ic" and "~ , ' ' ," and these terms wiii be used i..a,l.,ik llvvdW~ . These terms refer oniy to the primary structure of the 25 molecule. Thus, these terms include double- and single-stranded DNA, as weii as double-and single-stranded RNA amd DNA:RNA hybrids, amd aiso include known types of , for example, labels which are known in the art, methylation, "caps,"
substitution of one or more ofthe naturaily occurring nucleotides with an anaiog, inter-nucleotide " .... ~ . 6... ,~ such as, for example, those with uncharged linkages (e.g., methyl 30 1,l,,.~l,l,,~,, lr~ 1s ~ n' .~ ,carbamates,etc.)andwithchargedlinkages (e.g.,, ' , ' , ' ~Lua~llvlvd;llliùaLca, etc.), those containing pendant moieties, such as, for exarnple, proteins ~mcluding nucleases, toxins, antibodies, signai 2l968a6 096/06~04 9 : r~ 3~ 6 peptides, poly-~lysine, etc.), those with u~lL~Lu~ (e.g., acridine, psoralen, etc.), those containing chelators (e.g., metals, radioactive metals, boron, oxidative metals, etc.), those ~ containing alkylators, those with modified linkages (e.g., alpha anomeric nucleic acids, etc.), as well as unmodified forms ofthe polyl~uclcv~;dc or nl;g.. ~ ! vil~lf The term ",uu~ u~ ,vLIc analyte" or ",uvlyllucl~vL;df~ sample" refers to a single- or double-stranded nucleic acid molecule which contains a target nucleotide sequence. The analyte nucleic acids may be from a variety of sources, e.g., biological fluids or solids, food stuflfs, cll~;lull~ ,...dl materials, etc., and may be prepared for the hylJl;di~ iUII analysis by a variety of means, e.g., proteinase K/SDS, chaotropic salts, or the like. The term 10 "~ulJ...~ v~;L analyte" is used hllclu;~ge.lbly herein with the terms "analyte," "analyte nucleic acid," "target" and "target molecule." As used herein, the term "target region" or "target nucleotide sequence'' refers to a probe binding region contained within the target molecule. The term "target sequence" refers to a sequence with which a probe will form a stable hybrid under desired conditions.
It will be appreciated that, as used herein, the terms "uu~,k,06;de" and "lluclevt;dc"
will include those moieties which contain not only the known purine and pyrimidine bases, but also other L~,t~,.ul,y~ bases which have been modified. Such ~ include methylated purines or uy ~ " , acylated purines or pyrimidines, or other h.,t~,l u~,y~
Modified nucleosides or nucleotides will also include ~ on the sugar moiety, e.g., 2 0 wherein one or more of the hydro~syl groups are replaced with halogen, aliphatic groups, or are ~ " ,. l ;. ", l . 1 as ethers, amines, or the like.
As used herein, the term "probe" refers to a structure comprised of a poly..u~,lwl dc, as defined above, which contains a nucleic acid sequence ~ . ' y to a nucleic acid sequence present in the target molecule. The pol y l~u~.L,vL;Ic regions of probes may be 25 composed of DNA, and/or RNA, and/or synthetic nucleotide analogs.
The terms "nucleic acid multimer" or " ~ multimer" are used herein to refer to a linear or branched polymer of the same repeating single-stranded ~ vl;~le unit or different srngle-stranded puly~.ull~,vL;de units, each of which contains a region where a label probe can bind, i.e., contains a nucleic acid sequence ~ u"~l~l l A y to a nucleic acid 30 sequence contained within a label probe; the ~ vl; l~ units may be composed of RNA, DNA, modified nucleotides or r.~ thereo~ At least one of the units has a sequence, length, and, .. ,. ,.~ that permits it to bind specifically to a segment of a target WO 96/06l04 ~ 10 A ~,~ /i~ / /6 poly.l..~ ,vL;de; typically, such units will contain ~ y 15 to 50, preferably 15 to 30, nucleotides, and will have a GC content in the range of about 20% to about 80%. The total number of ~ ,I g~ f units in the multimer will usually be in the range of about 3 to 1000, more Iypically in the range of about 10 to 100, and most typicaily about 50. In one S type of bramched muitimer three or more ul;g, - ~ units emanate from a point of origin to fomm a bramched structure. The point of origin may be amother nucleotide unit or a molecule to which at least three units can be covaiently bound. In another type, there is an e 'i~ ' le unit backbone with one or more pendamt o~ g. . I~v~units linked to branch points in the backbone. These latter-type multimers are "fork-like,"
"comb-like" or ' "fork-" and "comb-like" in structure, wherein "comb-like"
muitimers are pu' ~ clcvLid~,~ having a linear backbone with a muitiplicity of sidechains extending from the backbone. Typically, there will be at least two branch points in the mUltimer~ more preferably at least three, more preferably in the range of about 5 to 30, although in some ~" l o~ there may be more. The multimer may include one or more1~ - lPV~ ; segments (e.g., comprised of protein nucleic acids or synthetic sequence-specific nucleic acid polymers, as noted above with respect to "~uly~ .lev~id~,~" in general), and one or more segments of double-stranded sequences. Further infommation concerning multimer synthesis and specific muitimer structures may be found in commoniy assigned U.S.
Patent No. 5,124,246 to Urdea et al.
2 o As used herein, a "biologicai sample" refers to a sample of tissue or fluid isolated from an individual, including but not limited to, for example, plasma, serum, spinal luid, semen, Iymph fluid, the extemal sections of the skin, respiratory, intestinal, and g y tracts, tears, saliva, milk, blood cells, tumors, orgams, amd also samples of in vilro cell culture constituents (including but not limited to conditioned medium resulting from the growth of cells in cell culture medium, putatively viraily infected cells, I~,ul~b;llrlllL celis, and cell components). Preferred uses of the present method are in detecting and/or q ~ virai antigens, such as from hepatitis B virus ("HBV"), hepatitis C virus ("HCV"), hepatitis D
virus ("HDV"), human ;., .... ~d 1~..;.... y virus ("HIV"), and the herpes family of viruses, includmg herpes zoster (chicken pox), herpes simplex virus I & Il, r;yi v;' .:.u~, Epstein-3 0 Barr virus, and the recently isolated Herpes VI virus.
By "protecting group" as used herein is meamt a species which prevents a segment of a molecule from undergoing a specific chemical reaction, but which is removable from the ..... .. , . ..... ..... . , ...... _ . . .. . . _ .. . _ ... . . ... ... . . _ . . , ... _ _ 2196~o~
~ WO 96/061~ PCT/US9S/10776 molecule following completion of that reaction. This is in contrast to a "capping group,"
which " I!! binds to a segment of a molecule to prevent any further chemical r. ", . -~ , 1 of that segment.
By "abasic site," as noted above, is meant a monomeric unit contained within a puly~ ,le~ide chain but which does not contain a purine or pyrimidine base. The term is used hlle~ herein with "modified dc.)~.ylil~ose residue". That is, the monomericunits used in conjunction with the method of the invention contain the dc~ y~ ibos~ ring but do not have a purine or pyrimidine base present at the I position.
The term "alkyl" as used herein refers to a branched or unbranched saturated h~ Ub-)ll group of I to 24 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, octyl, decyl, tetradecyl, hexadecyl, eicosyl, tetracosyl and the like.
Preferred alkyl groups herein contain I to 12 carbon atoms. The term "lower alkyl" intends an alkyl group of one to six carbon atoms, preferably one to four carbon atoms.
The term "alkylene" as used herein refers to a bifunctional saturated branched or unbranched l-yJ~ ,G.l,o-- chain containing from I to 24 carbon atoms, and includes, for example, methylene (-CH2-), ethylene (-CH2-CH2-), propylene (-CH2-CH2-CH2-), 2-methyl-propylene [-CH2-CH(CH3)-CH2-], hexylene [-(CH2)6-] and the like. "Lower alkylene" refers to an alkylene group of I to 6, more preferably I to 4, carbon atoms.
The term "aryl" as used herein refers to an aromatic species containing 1 to S
aromatic rings, either ",.~.,1,~61,.t. J or substituted with I or more substituents typically selected from the group consisting of-(CH2)~-NH2, -(CH2),~-COOH, -NO2, halogen and lower alkyl, where x is an integer in the range of 0 to 6 inclusive as outlined above. The term "aralkyl" intends a moiety containing both alkyl and aryl species, typically containing less than about 24 carbon atoms, and more typically less than about 12 carbon atoms in the alkyl segment of the moiety, and typically containing I to 5 aromatic rings. The term "aralkyl"
will usually be used to refer to aryl-substituted aikyl groups. The term "~lkyl,.l.~" will be used in a similar manner to refer to moieties containing both alkylene at~d aryl species, typically containing less than about 24 carbon atoms in the alkylene portion and I to 5 aromatic rings in the aryl portion, and typically aryl-substituted alkylene.
3 0 The term "arylene" refers to a difunctional aromatic moiety; "Il~v~lu~ , arylene"
refers to a phenylene group. These groups may be substituted with up to four ring substi-tuents as outlined above.
- 2~ Q~
WO96/06104 - 12- .._11~)~,3J~_//6 "Optional" or "optionaliy" means that the ~ubal;~u.,.lLly described event or uil~.ulllaL~ may or may not occur, and that the description includes instances where said event or ~,;., occurs and instances where it does not. For example, the phrase "optionally substituted alkylene" means that an alicylene moiety may or may not be 5 substituted and that the description includes both ~ alicylene and alicylene where there is 5nhctitntinn The Monomeric Reagents of the Invention:
The monomeric compounds of the invention which are used to create abasic sites 10 within pul~..u~,le~)L;.ie structures have the formula (I) (I) . ~, OR R
with R~, R2, R3, A, Z, X and n as defined above. It may be seen that reagent (I) is composed 2 o of a de~J,Lyl ;i,ose ring, containing substituents Rl and R2 at the 5 and 3 positions, respectively, which enable ;~U~u~,L;ull ofthe reagent into a pulylluclc~)L;de chain using coll~ ullol chemical DNA synthesis techniques. The moiety -A-Z-X(R9)~ at the I position replaces the purine or pyrtrnidine base normally present in a nucleotidic structure, and, as may be deduced from the def nition of R9, may be an u..~.,uL~Lci moiety, a protected 2 5 moiety, a labeled moiety, or a linker which is bound to a solid support.
Rl is, as noted above, a base-stable, acid-sensitive blocicing group. Such blocking groups are well-known in the art of..~ f synthesis and include u. ~ ~h~l;l..lr~ or substituted aryl or aralkyl groups, where the aryl is, e.g., phenyl, naphthyl, furanyl, biphenyl, or the liice, and where the substituents are from û to 3, usually to û to 2, and include any 3 0 ~ - I'rl ;1 Ig stable groups, neutral or polar, electron-donating or w;Lhll~w;ug. Examples ofsuchgroupsare.~ ,uAyLl;Lyl(DMT)~ b~yLI;Lyl(MMT)~tritylandpixyL A
particularly preferred moiety for use herein is DMT.
2l968~6 '' , wo 96/06104 - 13 - - r~ ,.,sl.~776 R2 is a phosphoriis derivative which is selècted so as to facilitate .,, 1 -0.~., of the reagent with the 5'-hydroxyl group of a nucleoside or an ~ IP chain. Such groups include I ' . ' ' ~ Jhù~iJhv~l k~s~ 7 pl~ , phosphites, H-~iJhuDiJllulull~oat~,~, and the like (see, e.g., EP Publication No. 0225807 by Urdea et al., 5 "Solution Phase Nucleic Acid Sandwich Assay and Poly"u~,L,uli(ie Probes Useful Therein,"
the disclosure of which is hl~,uliJul~lLr~d by reference herein.) Particularly preferred groups useful as R2 are iJhOaiJllul~L~IuJilt:~ having the structure:
N(iPr)~
1 0 _p~
O--Y
wherein Y is selected from the group consisîing of methyl and ~-cyanoethyl, and "iPr"
represents isopropyl. Most preferably, Y is ~3-cyanoethyl.
Aiternatively, R2 may be a linicage to a solid support, typically through a carbonyl moiety. That is, R2 may be -(CO)-RIc wherein Rl~ represents the solid support.
As noted above, the Rl and RZ substituents are generally selected so as to ailow;n~UI iJUI .l~;UII of the monomeric reagent (I) into a DNA fragment using standard r, chemistry protocols, well icnown in the art, and described, for example, in a2 0 number of the references cited hereinabove. In general, to incorporate the monomeric reagent (I) into a poly"u~ ide chain, the RZ substituent is selected so as enable reaction of the reagent at that position (i.e., the 3 position) with the 5'-hydroxyl group of a nucleoside or an ol ~ " 1 ul ;rl~ chain, while the Rl moiety is selected so as to enable reaction of the reagent at that position (i.e., the 5 position) with the 3'-hydroxyl of a nucleoside or an 25 ..li",.",.. 1. ~.~j.le chain.
Examples of preferred monomeric reagents r ~ L ~ d by structural formula (I) include the foiiowing:
WO96/06104 ~.96~~ .3,l "6 ~
--~R2 0-- --O' Z~ COCP3 R O
oR2 Z--N~
oR2 2l96Bo6 R/O~ NO z R10 O~S--S O~ 0 oR2 , S~
R10_~0 ~
oR2 .
R10 ~S--S ,~
-~ O
oR2 WO96/06104 Qo~6 r~ JU"6 ~,~9 1 6 -O
R10_, O~ ~ O--P--O-~ CH3 OH
oR2 ~lo O~s~
-~
R~O o~Si(CH3)3 ~Y
2s oR2 3 o O
~lo~ ~J ~C~H
3 5 oR2 2l968o6 ~ WO96/06104 17 PCT/US95/10776 S~O--~ t--RIO_~O~
RIO_~o~O~NH--CPG
oR2 Pul.yll..clcJL;;ie Reagents Containing Abasic Sites:
The ~-u'~ .-uclculide reagents of the invention which contain abasic sites are prepared using standard DNA synthesis chemistry and replacing a fraction of the nucleotidic monomers with n~ reagent (1). Generally, c~ w~ l.y I to 100% ofthe monomers used to synthesize the polynucleotide reagent will be replaced with reagent (1), more preferabiy 10 to 50~/0. and most preferably 20 to 40~/0 Generally, about 0 to 10 bases will be h~ul~ù~ e~l between n.~ r ~ n ~ monomer units. It is preferred, particularly when the R9 group is a large, bulky substituent, that the n~ t~ ' monomers (I) be spaced apart within the IJulyllu11.,vL;de chain. In such a case, at least about 3 bases should be ir.~u. ~)u. ~:Le i between monomer units to minimize steric interference or d ~ ~ -These pulr.~uuieuL;de reagents will generally have the structurai formuiae (Il), (111) or (IV) as shown above.
The polynucleotide reagents of the invention may be used as probes in a wide variety of hybridization assays such as those described in commonly assigned U.S. Patent Nos.
4,775,619 to Urdea et al., 4,868,105 to Horn et al., 5, i 18,605 to Urdea, 5, I Z4,246 to Urdea WO96/06104 ?~96~L6 - 18 - P~ 't~ 6 et al., 5,200,314 to Urdea~ as well as in PCT Publication Nos. 89/03891 (inventors Urdea et al.) and 92/22671 (inventors Horn et al.). Additionally, with respect to structures (Ill) and (IV), it shouid be noted that a 3'-3' linkage is provided, enabling use ofthe probes in triple helix formation.
In some cases, the linker arm present in n~ - - " ;~ monomer units resulting from ;n~ul~JuldLiUl~ of reagent (I) into the poly,-u~ vLide chain will contain a selectably cleavable site. Probes containing cleavable sites are particularly useful in the hyu,; i;LdliVII assay described in commonly assigned U.S. Patent No. 5,118,605 to Urdea et al., entitled 'Tuly..~l~,lcvL;de Determination with Selectable Cieavage Sites~" the disclosure of which is 10 ~ I,u-dLtvhereinbyreference, Thenatureofthecleavablesitemayvary,butwilltypically involve a linkage that may be cleaved using readily available chemical reagents, the only iimitation here being that the cleavage reagents are compatible with the various probes~
labels, etc., used in the remainder of the method. Generally, the cleavable site will be present in the moiety "Z" present within the R5 substituent in the formulae. Preferred cleavable sites arethoseidentifiedinU,S.PatentNo 5,118,605. Asexplainedinthatapplication,selectably cleavable sites include, for example, the ~;ullnwillg types of linkages:
O O
C ~ CH2 CH2 ~ (hydroxyla3nine-sensitive);
2û -C-NH- (b~se-sensitive);
O
-S- ( base-sens itive );
_5_5 _ ( thio1- s ens itive ); and Ol H Ol H
-CH--CH- ( periodate-sens itive ) .
N-hy h UAy ' ~ ' ~ (NHS) may be used to introduce the base-cleavable amide 30 bond into the reagent, while ethylene glycol bis(~ ~ ~ ~ 'yl succinate) may be used to create a hydroxyl~ , sen iitive linkage. bis[2-~ ' ~ yudl bùl~.~luAy)ethyl]sulfone (BSOCOES) may be used to create a ba~-sensitive sulfone linkage, ~ lyl tartarate 1 9 6 8 0 6 r, ~
~ WO 96/06104 P~ u / /6 (DST) may be used to introduce 1,2-diols cleavable by periodate, and dithiobis-~ 1~ ip, u~;~,.,atc) (DSP) may be used to provide thiol-cleavable disulfide bonds.
Methods of using these reagents to produce the desired cleavable linkage are well known and will be readily apparent to those skilled in the art of synthetic organic chemistry.
In the ar.,.c.. ,~.. ;u"cd ~ ~I.o~ , the moiety R9 represents a detectable label, such that cleavage of a linkage present within the spacer moiety Z will result in release of label .
Suitable labels which may be present at the R9 position in such a case include, for example, iu~ 4 ~, fluorescers, ' ' Imi~ srers~ dyes, enzymes, enzyme substrates, enzyme cofactors, enzyme inhibitors, enzyme subunits, metal ions, and the like. Illustrative specific 10 labels include fluorescein, rhodamine, Texas red, phycoerythrin, u".l,.llif~ , luminol, NADPH, a,b-~llr~iu~id~ .c, horseradish peroxidase, and the like.
Polynucleotide reagents useful as probes in hybridization assays may also be prepared by using the monomenc reagent (I) as a "branch poinl." In this way, probes containing branch points having the structural formula (V) - Ho r ~ Ai ~- ~ ~ P--~ ~ , ~, ,~ A ~ A ~ l-- G l~
O
1 ,5, ~
~r~--P--~-- ~ ~~11 o (V) wo96/06104 '1,~9 20- r~ lL ~/6 may be prepared, wherein DNA" DNA2, DNA3, R3, R4 and Rs are as defined above. Such 5 probes may be used, for example, in the ~ assays described in commoniy assigned U.S. Patent No. 5,124,246 to Urdea et al., entitied "Nucleic Acid Multimers and Amplified Nucleic Acid IIyblkl;~d~ivll Assays Using Same," PCT Publication Nos. W089/03891, and WO 92/02526. The latter application describes the comb-type branched multimers which are preferred in conjunction with the present method, and which are composed of a linear backbone and pendant sidechains; the backbone includes a segment that provides a specific hyl)li il~d~;UII site for anaiyte nucleic acid or nucleic acid bound to the analyte, whereas the pendant sidechains inciude iterations of a segment that provide specific Lrul;J;LdL;ull sites for a labeled probe.
In still another ~i.,Lo ihll~lL of the invention, the "abasic," or modified, site provided by monomeric reagent (I) may be used to enable synthesis of a ~ol~ .L.vliJu on a solid support. In this case, the reagent is bound to a solid support through the iinker arm at the I
position, i e., R9 represents a soiid support. As noted above, the linicage to the solid support may aiso be at the 3 position, at Rl Examples of solid supports include silica, Porasil C, polystyrene, controlled pore glass (CPG), kieselguhr, POIY(I~ GWYIG~Uid~
POIY(G~ llllUll ' ' ' ), polystyrene grafted onto pul.~ Lldlluulv~,Jl~ ;), cellulose, Sephadex LH-20 and Fractûsil 500. Nucleûtidic mûnomers are then added using standard DNA synthesis chemistry at the 3' and 5' positions. In some cases, i.e., to produce support-bound labeiied probes, it may be desirable to replace some nucleotidic monomers with labeiied monomers, e.g., the NJ-labelled cytidine derivatives described in commonly assigned U.S. Patent No. 5,093,232 to Urdea et al., entitled "Nucleic Acid Probes." Such monomers have the structurai formula (Vl) 2Ig680~ ' ~ WO96/06104 -21- r~ 1UI/6 R'~
R
(Vl) (~
1~
1 0 o~ ~ ~
~1~~o ~
OR
wherein R~ and Rl are as defined above;
R" is an optional linking moiety which, if present, contains an an~ide, thioether or disulfide linkage or a ~ .s,~ ;.." thereof;
2 o Rl2 is a reactive group d~,.iv~.Li~l,l,, with a detectable label, e.g., -NH~, -COOH or -SH;
R'3 is hydrogen, methyl, fluoro, bromo or iodo; and R'~ is either hydrogen, hydroxyl or protected hydroxyl.
In still another F~ .O- I : of the invention, I~UI ~ul,h,v~id~,3 are synthesized in which 25 themonomericreagent(I)maybeusedtochangethedirectionofsynthesis~e~g~from3~ 15'to 5' ~3' or vice versa. This is O ~ l by adding monomeric reagent (I) to the terminusofagrowingv~ vl;~lFchain~cappingeitherthe3~ors~terminalhydroxyl group with a capping group, typicaily an acyl capping group, and then using the l linker arm to continue synthesis in the reverse direction. Oligomers in which adjacent monomer units 3 0 are linked 3'-3' can also be prepared using reagent (I), by binding the ~ ;. IF to a solid support at R9, growing a single oligomer at the 5' position, capping exposed the WO 96106104 ~ 22 ~ J.,,J~ 6 group at the 5' terminus, and then growing a second oligomer at the 3' position. Such structures are illustrated in formulae (Il) and (III).
Synthetic Methods:
Scheme I illustrates the preferred method of ~yllLi. ,~;Lhlg monomeric reagents having the structural formula (1):
~ WO96/061~4 21968D6 p "~ o Scheme I
10 ~7~
o ' ~ ~
0~
9.' 1' ~ -~ ~
~r O7~ ~ v o ~ ¢7\~
o 1~ 7 S ,~ ~ 0 o ~5 , 30 ~s ~ X .1 ~!
o~
WO 96/06104 ?,~ 6 ~ / L l l6 In Scheme 1, 2-deoxy-D-, ;~ is used as the starting material. The three hydroxyl groups of the molecule are protected using an "R-CI" reagent or some other reagent suitable to protect free hydroxyl groups (e.g., benzoyl chloride or acetic anhydride) to provide 3 -OR groups at the I, 3 and 5 positions of the sugar. The product is isolated, and the l-OR group then replaced by reaction with a moiety (R9)~-X-~AH in the presence of an acid catalyst, followed by d.,~., uLeuL;on at the 3 and 5 positions using base. The S
position may then be selectively protected by reaction with Rl-CI, e.g., d;~ dlu~yLlilyl chloride, followed by reaction with a selected pl~ P at the 3 position to provide the desired phosphorus derivative.
The practice of the present invention will employ, unless otherwise indicated, ~,un~L;ull,~l techniques of synthetic organic chemistry, bio~,h~ L.y, molecular biology, and the like, which are within the skill of the art. Such techniques are explained fully in the literature. See, e.g., Sambrook, Fritsch & Maniatis, Molecular Clonin~ A J ol~nratnrv ~5~, Second Edition (1989); O8y~ ;de Svnthesis (M.J. Gait, ed., 1984), ~ç~ç~
Acid Ilvl ,. ;~ 1 (B.D. ~amcs & S.J. Higgins, eds.~ 1984); and a series, Methods in EI~YIIIOIUYY (Academic Press, Inc:). All patents, patent ~ , and I ' ' mentioned herein, both supra and i~u'ra, are hereby i~ ul~Jul~Ltd by reference.
2 0 It is to be understood that while the invention has been described in conjunction with the preferred specific ' ' thereof, that the description above as well as the exarnple which follows are intended to illustrate arLd not limit the scope of the invention. Other aspects, advantages and ,..n.l;i~ within the scope ofthe invention will be apparent to those skilled in the art to which the invention pertains.
In the following example, efforts have been made to insure accuracy with respect to numbers used (e.g., amounts, Ltl..~ LU.l:, etc.) but some c l~ ; ..;~1 error and deviation should be accounted for. Unless indicated otherwise, LtllllJ.,l~Lul~ is in degrees C and pressure is at or near ,a~ 8 2l968~6 .' ~',, ' ' ~ P~ 1 16 Exam~le I
1,3,5-Tris-O-acyl-2-deoxy-D-,;i,uru,~,lu~c was readily synthesized by treating CU~ILI.~ Y available 2-deoxy-D-. ;1,. ,~: " .., ~. .~ with a large excess of acetic anhydride or benzoyl chloride in pyridine. Both 1,3,5-O-trisacetyi- and trisbenzoyl-2-deoxy-5 D-,;i,uru"..oaG couid be ~ G~l y ' "' ~ firom ethanol. The acetyl derivative was mainiy the aipha-isomer, and the benzoyl derivative gave the two isomers in 1/1 ratio. No pyrancside derivative was formed ( less than 5% ).
1,3,5-O-tris(TBDMS)-2'-deoxy-D-, ;1 ", ~ 5itlr- was synthesized from ieU~Lyl ;iJo:~G by reaction with t-buLyl ihl.~..llybil~l ~,1 iu,;dc';"..d, ~JIc/DMF.
The anomeric acetai was readily exchanged with an alcohol in the presence of an acid catalyst, such as ZnBr2, to give the alcohol derivative of either 3,5-0-diacyl- or 3,5-O- di-TBDMS -2'-deoxy-, ;1 " ,1; ", ~f Removal of the 3,5-O-protecting groups with base (methanoi/lM K2CO3 for acyi) or fluoride ions (IM LGLI.LI,UIYI~"U,-U....I-., fluoride in THF
for TBDMS) gave the substituted 2'-deoxy- ~;burul~ uac derivatives.
Aicoholscontainingvarious li, li.~ ; havebeen;.. ~,u,pu,dltdthisway.
R~,yl~"c~t~L;vG examples are 4-1ll~,.~.ylu~y-".-lJu--yllJ.,.~yl, 4-1ut~uyll~.l..,Lllyl~
TFA-NH-alicyl(aryl), and N-(4~u~ ub~ Iu~y~;~uul~yl)/
~;~IOC-6-aminohexyl. They were aii prepared directly from the CUIlGyUlld;ll~; alcohol and 1,3,5-tri-0-acyl-2'-deoxy-D-,ii,uru..l,,uaG.
2 0 Preparation of S-trityl- I l-mercapto- I-undecyl was achieved via the 11 -bromo-l-undecyl derivative: After preparation of the 11 -bromo- I -undecyl 3,5-di-O-acetyl-2-deoxy-D-,ii,uL~l~u,uaG reaction with ~iLyh~,.l , (Tr-SH) in the presence of base (one equivaient of aq. NaOH ) afforded S-trityl- 11-u~ yLuulld~,~.yl 2-deoxy-D-.;l,~ r~ Aiternatively, S-Tr-ll-mercapto-l-undecanol could be prepared and used as the aicohol component. Aiternatively, it is possible to incorporate aicohols containing a disulfite, -S-S-. The O-levuiinyl-l l-oxo-undecyl derivative was prepared via the 11 -bromo derivative. After removai of the acetyl ~roups, of bromine with the Cs-salt of levulinic acid afforded O-levulinyl-l l -oxy-undecyl 2-deoxy-D-,ii,, ~ ~ Aiternatively, preformed O-levulinyl-l l-oxy-l-undecanol 3 0 could be used as the alcohol component.
The appropriate alicyl 2-deoxy-D-~ il,uru- a"uaidG analogs were converted to theS-DMT derivatives using stamdard literature procedures. The two anomeric aklGu;Su..._l a ,, . .. . ...... ... . _ WO 96/06104 ?~ 26 P~ 16 gave rise to DMT species with quite different mobilities during silica gel clll~ " . ', .
All DMT ' were purified by silica gel ~,LI ulllaLu~ lly, and the two anomeric ~Lcl CU;~UIII~ were readily separated. The various DMT ;"~ were converted to the 3-0-N,N-d;;~u~luuyl~.yGIlu~,;llyl-~ lln~ using standard literature procedures, and they could be used like normal nucleoside uy.-llu~ yl~ during automated synthesis.
Removal of protecting groups from chemically synthesized 1;". ~ ul; '~ ~ required only minimal changes to the standard procedures.
~ ..,.,,h~ !u~y~,albu~l ~ lu~l~yl 2-deoxy-D-~ u~ Hydrolysis of methyl ester and succinate linkage to support was carried out with water/TEA/dioxane (I :1:10 v/v; 13 hours) prior to exposure to ammonium hydroxide.
TFA/FMOC-NH-alkyl required only standard deprotection with ammonium hydroxide. For 4-1l;LIu~L~ ;llyl and N-(4--~ ub.,.l~lu~y-carbonyl)-6-~lli-lùli~yl.
reduction of the nitro group to an anilino group was conducted with 0.1 M sodium15 dithionitellM TEAB/ dioxane for S hours, washed, and then deprotected with ammonium hydroxide to give the free anilino- and amino derivatized oligomer, l~ ,.,Li~ , which was purified by PAGE.
On support treatment with HPAA reagent, the DNA synthesis can be continued on the same support to produce branched oligomers. With proper choice of side-arm length, 2 0 the monomer is usefiul for making 3'-3' linked oli~,uu~ uLid~ for cross-over triple helix formation. An exarnple is O-levulinyl-2-oxyethyl S-DMT-0-2-deoxy-D-, il ,. .ll .. ~
3'-O-succ-CPG; the first strand is synthesized using 5'-DMT, capped, the levulinyl group removed and synthesis continued at the 2-h~d~u~ ;h~l side-chain. Deprotection gives the desired 5'-DNAI-3'3'-DNA2-5' oligomer.
Thus, in addition to utility in providing cleavable sites wjthin, 'i~ ' ' or l)ul~..u~ ,v~ide chains, the invention enables a number of procedures deriving from the presence of linker arms at the I position of a monomeric dcv~yl ib~/a~. unit rather than purine or pyrimidine bases as present in cu~ liu~ nucleotide structures.
2 0 Overview of thP Art Backgrûund references which relate generally to methods for bylllh~;~
.,1;5 """ l~vl~' include those related to 5'-to-3' syntheses based on the use of,B-cyanoethyl phosphate protecting groups, e.g., de Napoli et al., Gazz Chim rt~l 114:65 (1984), Rosenthal et aL, Tetrrhp~rûn T .PttPrS 24: 1691 (1983),1Belagaje and Brush, Nucleic ~ri~lc F PCP~rrh 10:6295 (1977), in references which describe solution-phase 5'-to-3' syntheses include Hayatsu and Khorana, J American ~'hPmir~l Society 89:3880 (1957), Gait amd Sheppard, ~llrlPir Aririe RP~P~rrh 4 1135 (1977), Cramer and Koster, Aneew ('hPm Int E.l Fn~d _:473 (1968), and Blackbum et al., Journal ofthe Chemical Society, Part C, 2438 (1967).
In addition to the above-cited art, Matteucci and Caruthers, J. AmPrir~n ~hP.n;. ~1 Society 103:3185-3191 (1981), describe the use of ~ .h. ~ in the preparation of r~ 1 vl; !~ ~ Beaucage and Caruthers, Tetrahedron Letters;~:1859-1862 (1981), and U S Patent No 4~415~732 describe the use of l l ~ in the preparation of . _, .. , ., .. , .. . _ , _ , , 2~96806 '~ r s ~ WO96/06104 3 PCT/US95/10776 . vL; !r~ Smith, ABL 15-24 (December 1983), describes automated solid-phase o:;gvJ~v~yl il ;-, ,. .1, . .~h;P synthesis. See also the references cited therein, and Warner et al., DNA 3 :401411 (1984), whose disclosure is h~,u~5~u~ J herein by reference.
U.S. Patent Nos. 4,483,964 and 4,517,338 to Urdea et al. describes a method for S ~y..~h.,~ g pol~"...,l~,vfid~,s by selectively introducing reagents to a solid phase substrate in a tubular reaction zone. U.S. Patent No. 4,910,300 to Horn et al. also describes a method for ~y~lih.,~ g rl;r","." If . ,I;s~ - by sequentially adding nucleotidic monomers to a growing chain, but involves the in~ùl ~u~ ~L;ull of labelled, N-4 modified cytosine residues at ~-~d~ ... J, spaced apart positions. U.S. Patent No. 5,256,549 to Horn et al. is also of 1 û interest in that a method for preparing .~ f vl;: - is provided which involves a -6. .,. technique, i.e., in which the desired ,~ . ,"",.1. v~ is essentially synthesized and "purified" ~ f. ~ ly~ such that the final product is produced in ~ , pure form.
HomandUrdea,DNA5(5):421-425(1986),describe~uhv~vhvlyl~Liullofsolid-15 supported DNA fragments using bis(~ ..u~il.w~y)-N,N-diisopropyl ,t ~ See also, Horn and Urdea, Tetrahedron T PtfPr~ 27:47054708 (1986).
References which relate to hybl;J;~L;ùn techniques in general include the following:
Meinkoth and Wahl, Anal. Biol '...,. ny 138:267-284 (1984), provide an excellent review of hys,l;J;~L;u~ techniques. Leary et al., Proc. Natl. Ar Irl Sci. ~fUSA) 80:4045-4049 (1983) 2 0 describe the use of biotinylated DNA in conjunction with an avidin-enzyme conjugate for detectionofspecifico!;,~v,,lcl~.vL;Jesequences. R~nkietal.,Gene21:77-85,describewhat they refer to as a "sandwich" hylJI;J;~L;ull for detection of, .I;g. ,~ sequences.
Pfeuffer and Helmrich, J. Biol. Chem. ~:867-876 (1975), describe the coupling ofguanosine-5'-0-(3-i'i . ' - . ' ) to Sepharose 4B. Bauman et al., J. Hi~fr r hPm ~n~
Cvtochem 29:227-237, describe the 3'-labeling of RNA with fluorescers. PCT Application 7 describes the addition to DNA fragments of modified l;b.. If vl ;. l. for labeGng and methods for analyzing such DNA fragments. Renz and Kurz, Nucl. Ari~c Rr~
12:3435-3444, describe the covalent linking of enzymes to u~ ; ' - Wallace, DNA
R~ ~ T~ Woo, S., ed.) CRC Press, Boca Raton, Florida, provides a 3 0 general background of the use of probes in diagnosis. Chou and Merigan, N. ~;n,p J of ~L 308 921-925, describe the use of a ~ JI~5~e-labeled probe for the detection of CMV. Lnm3r~ M,-fhr,l~ in Fn7vmol 34B, 24:77-102 (1974), describes procedures for Wos6/06io4 &~S6 ~ //6 linking to poly~,.,.yhu~l;d~,." while Parikh et al., Methods in Enzymol. 34B, 24:77-102 (1974) describe coupling reactions with agarose. Aiwine et al., Proc. Natl. Acad. Sci. (USA) 74:5350-5354 (1977), describe a method of transferring .~ from gels to a solid support for hyblici;LGl;ull. Chu et al., Proc. Natl. Acad. Sci. (USA) 11 :6513-6529, describe a technique for derivatizing temlinal nucleotides. Ho et ai., F~ . .h. . . ,;~l ~y 20:64-67 (1981), describe derivatizing temlinal nucleotides through phosphate to fomm esters. Ashiey and MacDonaid, Anai. Bionh~m 140.95-103 (1984), report a method for preparing probes from a surface-bound template.
Home and Dervan, J. Am. Chem. Soc I i2:2435-2437 (1990), and Froehier et ai., Bi(lrh~mi~hv 31: 1603- 1 ~)9 (1992), relate to . .1;.~ r vl ;- Ie-directed triple helix formation.
Summarv of the Inv~ nti~ln in one aspect of the invention, then7 monomeric reagents useful for providing the 15 novel polyllu~levL;de structures are provided, the monomeric reagents having the structural fommula (I) ~ R4 ~ ~Rs wherein:
R' is selected from the group consisting of hydrogen, acid-sensitive, base-stable protecting groups and acyl capping groups;
R2 is a phosphorus derivative selected to enable addition of the reagent to a 30 molecular species containing a free hydroxyl group, or is a linkage to a solid support;
R3 is selected from the group consisting of hydrogen, hydroxyl, sulfhydryl, halogeno, amino, aikyl, allyl~ oR6 wherein R6 is alkyl, allyl, silyl or phosphate;
~ WO96/06104 19680~ s ' r~ 6 R4 is either hydro~en or -(CH2)mOR7 wherein R' is alicyl or -(Co)R5, R5 is alL-yl, and m is an integer in the range of 0 to 12 inclusive;
Rs is ~A~Z~X(R9)n;
A is oxygen, sulfur or methylene;
~ 5 Z is arylene, C6-C,5 aralicylene or C~-C~z alkylene containing 0 to 6 I.~,Ltlu~.Lu,.. .
selected from the group consisting of O, S, N, Si and Se and 0 to 6 linL-ages selected from the group consisting of -CO-,-COO-, -CONH-, -NHCO-, -S-S-, -SOz-, -CH(OH)-CH(OH)-, -CH(oR4)-CH(oR4)-, -O-PO(O~-O-, -o-Po(R4)-,-o-Po(oR4)-o-7 -o-Po(oR4)-R5- and -Po(oR4)-o-R5- in which R4 is lower aiLyl and R5 is lower alLylene, and, if Z is aralLylene or 10 allylene, containing 0 to 3 unsaturated bonds;
X is selected from the group consisting of-NH-, -CONH-, -NHCO-, -CO-, -S- and -SiE;
R9 is hydrogen, a protecting group, a detectable label, or, uniess X is _SjE, a solid support; and n is I when X is -NH-, -CONH-, -NHCO-, -CO-, or -S-, and is 3 when X is -Si=.
In another aspect, ~oir..,~ ,Li i~ reagents are provided having the structurai formulae (II), (III) or (IV) ~ ~lo ~ o--P--" ~ R
Rs . O R~
2 5 11 o--P--o-- ~ ~ to A 1--~
a r ~9~Q6 WO 96106104 - 6 - - - r~ O
s ~C--p_ ~ 4 (111) O~ R
5'- ~ c-&~o~ 4 ~1~ \~p~
(IV) ~ P-O -~ IDN~ ~S-C~
wherein DNA, represents a first segment of DNA DNA~ represents a second segment of DNA and R3, R~ and R5 are as defined above. In a related aspect of the invention, branched DNA is provided having the structural formula (V) WO96/06104 9680~ 7 r~l~u..,J,i l16 1~~ ~hlA~ ~- c - ~- O ~
~ CA ~ a l'l A ~ 1-- ~ l~
~ ~ lP o ~ o ~.~
10 wherein DNA,, DNA2 and DNA3 represent firso second and third segments of DNA, and R;, R', A and Z are as defined above.
In still other aspects of the invention, methods are provided for synthesizing pcl~ iJ~ containing abasic sites and for preparing branched DNA. These methods involve the ;-,~u- ~,u, 4L;UII of the above-mentioned monomeric reagent into larger 15 p~ ..u~,L,uL;de structures.
A method is also provided for detecting the presence of an ~I g- ~ u~ e sequenceof interest in a sample which involves hybridizing the nucleic acid sample with a pGlJ"~,leolide probe containing an abasic site as described herein, wherein the abasic site is formed from the monomeric reagent defined above, and fiurther wherein the reagent contains 2 0 a detectable label at R3 and a cleavable site within the linker moiety -Z-. Either the sample or the po .~ ,vLide probe is bound to a solid support, such that hybridization results in a label being bound to the support through the cleavable site. Following l,yb, ;.1;~4fio-" the cleavable site is cleaved with a suitable reagent so as to release the detectable label R3, and label which is free of the support is quantitated and correlated with the presence andlor quantity of 2 5 sample.
Additionally, probes synthesized using the compounds of the invention may contain 3'-3' linkages, as illustrated in structures (Ill) and (IV) above. Ol;6oue~J~.yllu~l~ul;ll~ probes containing 3'-3' linkages can be used in triple helix formation, i.e., as such probes can bind to opposite strands of duplex DNA.
WO96/06104 2~ 96~~6 - 8i - P~ /6 1 pPt~ i Descrivtion of the Invention Definitions and .~. ," .. ,1 l ". ,:
Before the present invention is disclosed and described in detail, it is to be understood that this invention is not iimited to specific assay formats, materials or reagents, 5 as such may, of course, vary. It is aiso to be understood that the i ~ vJ used herein is for the purpose of describing particular r ~1 ~0/~ oniy and is not intended to be limiting.
It must be noted that, as used in the ~ ;..,. amd the appended claims, the singular forms "a," "an" and "the" include plurai referents uniess the context clearly dictates otherwise. Thus, for example, reference to "a monomeric reagent" includes mixtures of 10 monomeric reagents, reference to "a pu4..l~,1evL;de probe" may include mixtures of different probes, reference to a po4..~1vlLvLiJc containing "an abasic site" includes l~u4~u~,lvvLi;lC~
containing two or more abasic sites, and the like.
In this ~ .", and in the claims which follow, reference will be made to a number of terms which shail be defined to have the following meanings:
As used herein, the terms "~u4.~ evLi ie'~ and ". 1~ " shail be generic to pcl~J~vAyl ;1~ L ..1;.5. ~ (containing 2-deoxy-D-ribose), to ~uly. ;1..., . 1 .1; 1. (containing D-ribose), to any other type of po4..v~ Iev~ide which is an N-giycoside of a purine or pyrimid;ne base, and to other polymers containing ~ backbones (e.g., protein nucleic acids and synthetic sequence-specific nucleic acid polymers vUIIIII~ available 2 0 from the Anti-Gene Development Group, Corvailis, Oregon, as NeugeneTM polymers), providing that the polymers contain .,. ,. 1~ ob ~-~ in a ~ 5v ~iun which ailows for base pairing and base stacking, such as is found in DNA and RNA. There is no intendeddistinction in length between the term "I,c~l~.lJ~ .vLi ic" and "~ , ' ' ," and these terms wiii be used i..a,l.,ik llvvdW~ . These terms refer oniy to the primary structure of the 25 molecule. Thus, these terms include double- and single-stranded DNA, as weii as double-and single-stranded RNA amd DNA:RNA hybrids, amd aiso include known types of , for example, labels which are known in the art, methylation, "caps,"
substitution of one or more ofthe naturaily occurring nucleotides with an anaiog, inter-nucleotide " .... ~ . 6... ,~ such as, for example, those with uncharged linkages (e.g., methyl 30 1,l,,.~l,l,,~,, lr~ 1s ~ n' .~ ,carbamates,etc.)andwithchargedlinkages (e.g.,, ' , ' , ' ~Lua~llvlvd;llliùaLca, etc.), those containing pendant moieties, such as, for exarnple, proteins ~mcluding nucleases, toxins, antibodies, signai 2l968a6 096/06~04 9 : r~ 3~ 6 peptides, poly-~lysine, etc.), those with u~lL~Lu~ (e.g., acridine, psoralen, etc.), those containing chelators (e.g., metals, radioactive metals, boron, oxidative metals, etc.), those ~ containing alkylators, those with modified linkages (e.g., alpha anomeric nucleic acids, etc.), as well as unmodified forms ofthe polyl~uclcv~;dc or nl;g.. ~ ! vil~lf The term ",uu~ u~ ,vLIc analyte" or ",uvlyllucl~vL;df~ sample" refers to a single- or double-stranded nucleic acid molecule which contains a target nucleotide sequence. The analyte nucleic acids may be from a variety of sources, e.g., biological fluids or solids, food stuflfs, cll~;lull~ ,...dl materials, etc., and may be prepared for the hylJl;di~ iUII analysis by a variety of means, e.g., proteinase K/SDS, chaotropic salts, or the like. The term 10 "~ulJ...~ v~;L analyte" is used hllclu;~ge.lbly herein with the terms "analyte," "analyte nucleic acid," "target" and "target molecule." As used herein, the term "target region" or "target nucleotide sequence'' refers to a probe binding region contained within the target molecule. The term "target sequence" refers to a sequence with which a probe will form a stable hybrid under desired conditions.
It will be appreciated that, as used herein, the terms "uu~,k,06;de" and "lluclevt;dc"
will include those moieties which contain not only the known purine and pyrimidine bases, but also other L~,t~,.ul,y~ bases which have been modified. Such ~ include methylated purines or uy ~ " , acylated purines or pyrimidines, or other h.,t~,l u~,y~
Modified nucleosides or nucleotides will also include ~ on the sugar moiety, e.g., 2 0 wherein one or more of the hydro~syl groups are replaced with halogen, aliphatic groups, or are ~ " ,. l ;. ", l . 1 as ethers, amines, or the like.
As used herein, the term "probe" refers to a structure comprised of a poly..u~,lwl dc, as defined above, which contains a nucleic acid sequence ~ . ' y to a nucleic acid sequence present in the target molecule. The pol y l~u~.L,vL;Ic regions of probes may be 25 composed of DNA, and/or RNA, and/or synthetic nucleotide analogs.
The terms "nucleic acid multimer" or " ~ multimer" are used herein to refer to a linear or branched polymer of the same repeating single-stranded ~ vl;~le unit or different srngle-stranded puly~.ull~,vL;de units, each of which contains a region where a label probe can bind, i.e., contains a nucleic acid sequence ~ u"~l~l l A y to a nucleic acid 30 sequence contained within a label probe; the ~ vl; l~ units may be composed of RNA, DNA, modified nucleotides or r.~ thereo~ At least one of the units has a sequence, length, and, .. ,. ,.~ that permits it to bind specifically to a segment of a target WO 96/06l04 ~ 10 A ~,~ /i~ / /6 poly.l..~ ,vL;de; typically, such units will contain ~ y 15 to 50, preferably 15 to 30, nucleotides, and will have a GC content in the range of about 20% to about 80%. The total number of ~ ,I g~ f units in the multimer will usually be in the range of about 3 to 1000, more Iypically in the range of about 10 to 100, and most typicaily about 50. In one S type of bramched muitimer three or more ul;g, - ~ units emanate from a point of origin to fomm a bramched structure. The point of origin may be amother nucleotide unit or a molecule to which at least three units can be covaiently bound. In another type, there is an e 'i~ ' le unit backbone with one or more pendamt o~ g. . I~v~units linked to branch points in the backbone. These latter-type multimers are "fork-like,"
"comb-like" or ' "fork-" and "comb-like" in structure, wherein "comb-like"
muitimers are pu' ~ clcvLid~,~ having a linear backbone with a muitiplicity of sidechains extending from the backbone. Typically, there will be at least two branch points in the mUltimer~ more preferably at least three, more preferably in the range of about 5 to 30, although in some ~" l o~ there may be more. The multimer may include one or more1~ - lPV~ ; segments (e.g., comprised of protein nucleic acids or synthetic sequence-specific nucleic acid polymers, as noted above with respect to "~uly~ .lev~id~,~" in general), and one or more segments of double-stranded sequences. Further infommation concerning multimer synthesis and specific muitimer structures may be found in commoniy assigned U.S.
Patent No. 5,124,246 to Urdea et al.
2 o As used herein, a "biologicai sample" refers to a sample of tissue or fluid isolated from an individual, including but not limited to, for example, plasma, serum, spinal luid, semen, Iymph fluid, the extemal sections of the skin, respiratory, intestinal, and g y tracts, tears, saliva, milk, blood cells, tumors, orgams, amd also samples of in vilro cell culture constituents (including but not limited to conditioned medium resulting from the growth of cells in cell culture medium, putatively viraily infected cells, I~,ul~b;llrlllL celis, and cell components). Preferred uses of the present method are in detecting and/or q ~ virai antigens, such as from hepatitis B virus ("HBV"), hepatitis C virus ("HCV"), hepatitis D
virus ("HDV"), human ;., .... ~d 1~..;.... y virus ("HIV"), and the herpes family of viruses, includmg herpes zoster (chicken pox), herpes simplex virus I & Il, r;yi v;' .:.u~, Epstein-3 0 Barr virus, and the recently isolated Herpes VI virus.
By "protecting group" as used herein is meamt a species which prevents a segment of a molecule from undergoing a specific chemical reaction, but which is removable from the ..... .. , . ..... ..... . , ...... _ . . .. . . _ .. . _ ... . . ... ... . . _ . . , ... _ _ 2196~o~
~ WO 96/061~ PCT/US9S/10776 molecule following completion of that reaction. This is in contrast to a "capping group,"
which " I!! binds to a segment of a molecule to prevent any further chemical r. ", . -~ , 1 of that segment.
By "abasic site," as noted above, is meant a monomeric unit contained within a puly~ ,le~ide chain but which does not contain a purine or pyrimidine base. The term is used hlle~ herein with "modified dc.)~.ylil~ose residue". That is, the monomericunits used in conjunction with the method of the invention contain the dc~ y~ ibos~ ring but do not have a purine or pyrimidine base present at the I position.
The term "alkyl" as used herein refers to a branched or unbranched saturated h~ Ub-)ll group of I to 24 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, octyl, decyl, tetradecyl, hexadecyl, eicosyl, tetracosyl and the like.
Preferred alkyl groups herein contain I to 12 carbon atoms. The term "lower alkyl" intends an alkyl group of one to six carbon atoms, preferably one to four carbon atoms.
The term "alkylene" as used herein refers to a bifunctional saturated branched or unbranched l-yJ~ ,G.l,o-- chain containing from I to 24 carbon atoms, and includes, for example, methylene (-CH2-), ethylene (-CH2-CH2-), propylene (-CH2-CH2-CH2-), 2-methyl-propylene [-CH2-CH(CH3)-CH2-], hexylene [-(CH2)6-] and the like. "Lower alkylene" refers to an alkylene group of I to 6, more preferably I to 4, carbon atoms.
The term "aryl" as used herein refers to an aromatic species containing 1 to S
aromatic rings, either ",.~.,1,~61,.t. J or substituted with I or more substituents typically selected from the group consisting of-(CH2)~-NH2, -(CH2),~-COOH, -NO2, halogen and lower alkyl, where x is an integer in the range of 0 to 6 inclusive as outlined above. The term "aralkyl" intends a moiety containing both alkyl and aryl species, typically containing less than about 24 carbon atoms, and more typically less than about 12 carbon atoms in the alkyl segment of the moiety, and typically containing I to 5 aromatic rings. The term "aralkyl"
will usually be used to refer to aryl-substituted aikyl groups. The term "~lkyl,.l.~" will be used in a similar manner to refer to moieties containing both alkylene at~d aryl species, typically containing less than about 24 carbon atoms in the alkylene portion and I to 5 aromatic rings in the aryl portion, and typically aryl-substituted alkylene.
3 0 The term "arylene" refers to a difunctional aromatic moiety; "Il~v~lu~ , arylene"
refers to a phenylene group. These groups may be substituted with up to four ring substi-tuents as outlined above.
- 2~ Q~
WO96/06104 - 12- .._11~)~,3J~_//6 "Optional" or "optionaliy" means that the ~ubal;~u.,.lLly described event or uil~.ulllaL~ may or may not occur, and that the description includes instances where said event or ~,;., occurs and instances where it does not. For example, the phrase "optionally substituted alkylene" means that an alicylene moiety may or may not be 5 substituted and that the description includes both ~ alicylene and alicylene where there is 5nhctitntinn The Monomeric Reagents of the Invention:
The monomeric compounds of the invention which are used to create abasic sites 10 within pul~..u~,le~)L;.ie structures have the formula (I) (I) . ~, OR R
with R~, R2, R3, A, Z, X and n as defined above. It may be seen that reagent (I) is composed 2 o of a de~J,Lyl ;i,ose ring, containing substituents Rl and R2 at the 5 and 3 positions, respectively, which enable ;~U~u~,L;ull ofthe reagent into a pulylluclc~)L;de chain using coll~ ullol chemical DNA synthesis techniques. The moiety -A-Z-X(R9)~ at the I position replaces the purine or pyrtrnidine base normally present in a nucleotidic structure, and, as may be deduced from the def nition of R9, may be an u..~.,uL~Lci moiety, a protected 2 5 moiety, a labeled moiety, or a linker which is bound to a solid support.
Rl is, as noted above, a base-stable, acid-sensitive blocicing group. Such blocking groups are well-known in the art of..~ f synthesis and include u. ~ ~h~l;l..lr~ or substituted aryl or aralkyl groups, where the aryl is, e.g., phenyl, naphthyl, furanyl, biphenyl, or the liice, and where the substituents are from û to 3, usually to û to 2, and include any 3 0 ~ - I'rl ;1 Ig stable groups, neutral or polar, electron-donating or w;Lhll~w;ug. Examples ofsuchgroupsare.~ ,uAyLl;Lyl(DMT)~ b~yLI;Lyl(MMT)~tritylandpixyL A
particularly preferred moiety for use herein is DMT.
2l968~6 '' , wo 96/06104 - 13 - - r~ ,.,sl.~776 R2 is a phosphoriis derivative which is selècted so as to facilitate .,, 1 -0.~., of the reagent with the 5'-hydroxyl group of a nucleoside or an ~ IP chain. Such groups include I ' . ' ' ~ Jhù~iJhv~l k~s~ 7 pl~ , phosphites, H-~iJhuDiJllulull~oat~,~, and the like (see, e.g., EP Publication No. 0225807 by Urdea et al., 5 "Solution Phase Nucleic Acid Sandwich Assay and Poly"u~,L,uli(ie Probes Useful Therein,"
the disclosure of which is hl~,uliJul~lLr~d by reference herein.) Particularly preferred groups useful as R2 are iJhOaiJllul~L~IuJilt:~ having the structure:
N(iPr)~
1 0 _p~
O--Y
wherein Y is selected from the group consisîing of methyl and ~-cyanoethyl, and "iPr"
represents isopropyl. Most preferably, Y is ~3-cyanoethyl.
Aiternatively, R2 may be a linicage to a solid support, typically through a carbonyl moiety. That is, R2 may be -(CO)-RIc wherein Rl~ represents the solid support.
As noted above, the Rl and RZ substituents are generally selected so as to ailow;n~UI iJUI .l~;UII of the monomeric reagent (I) into a DNA fragment using standard r, chemistry protocols, well icnown in the art, and described, for example, in a2 0 number of the references cited hereinabove. In general, to incorporate the monomeric reagent (I) into a poly"u~ ide chain, the RZ substituent is selected so as enable reaction of the reagent at that position (i.e., the 3 position) with the 5'-hydroxyl group of a nucleoside or an ol ~ " 1 ul ;rl~ chain, while the Rl moiety is selected so as to enable reaction of the reagent at that position (i.e., the 5 position) with the 3'-hydroxyl of a nucleoside or an 25 ..li",.",.. 1. ~.~j.le chain.
Examples of preferred monomeric reagents r ~ L ~ d by structural formula (I) include the foiiowing:
WO96/06104 ~.96~~ .3,l "6 ~
--~R2 0-- --O' Z~ COCP3 R O
oR2 Z--N~
oR2 2l96Bo6 R/O~ NO z R10 O~S--S O~ 0 oR2 , S~
R10_~0 ~
oR2 .
R10 ~S--S ,~
-~ O
oR2 WO96/06104 Qo~6 r~ JU"6 ~,~9 1 6 -O
R10_, O~ ~ O--P--O-~ CH3 OH
oR2 ~lo O~s~
-~
R~O o~Si(CH3)3 ~Y
2s oR2 3 o O
~lo~ ~J ~C~H
3 5 oR2 2l968o6 ~ WO96/06104 17 PCT/US95/10776 S~O--~ t--RIO_~O~
RIO_~o~O~NH--CPG
oR2 Pul.yll..clcJL;;ie Reagents Containing Abasic Sites:
The ~-u'~ .-uclculide reagents of the invention which contain abasic sites are prepared using standard DNA synthesis chemistry and replacing a fraction of the nucleotidic monomers with n~ reagent (1). Generally, c~ w~ l.y I to 100% ofthe monomers used to synthesize the polynucleotide reagent will be replaced with reagent (1), more preferabiy 10 to 50~/0. and most preferably 20 to 40~/0 Generally, about 0 to 10 bases will be h~ul~ù~ e~l between n.~ r ~ n ~ monomer units. It is preferred, particularly when the R9 group is a large, bulky substituent, that the n~ t~ ' monomers (I) be spaced apart within the IJulyllu11.,vL;de chain. In such a case, at least about 3 bases should be ir.~u. ~)u. ~:Le i between monomer units to minimize steric interference or d ~ ~ -These pulr.~uuieuL;de reagents will generally have the structurai formuiae (Il), (111) or (IV) as shown above.
The polynucleotide reagents of the invention may be used as probes in a wide variety of hybridization assays such as those described in commonly assigned U.S. Patent Nos.
4,775,619 to Urdea et al., 4,868,105 to Horn et al., 5, i 18,605 to Urdea, 5, I Z4,246 to Urdea WO96/06104 ?~96~L6 - 18 - P~ 't~ 6 et al., 5,200,314 to Urdea~ as well as in PCT Publication Nos. 89/03891 (inventors Urdea et al.) and 92/22671 (inventors Horn et al.). Additionally, with respect to structures (Ill) and (IV), it shouid be noted that a 3'-3' linkage is provided, enabling use ofthe probes in triple helix formation.
In some cases, the linker arm present in n~ - - " ;~ monomer units resulting from ;n~ul~JuldLiUl~ of reagent (I) into the poly,-u~ vLide chain will contain a selectably cleavable site. Probes containing cleavable sites are particularly useful in the hyu,; i;LdliVII assay described in commonly assigned U.S. Patent No. 5,118,605 to Urdea et al., entitled 'Tuly..~l~,lcvL;de Determination with Selectable Cieavage Sites~" the disclosure of which is 10 ~ I,u-dLtvhereinbyreference, Thenatureofthecleavablesitemayvary,butwilltypically involve a linkage that may be cleaved using readily available chemical reagents, the only iimitation here being that the cleavage reagents are compatible with the various probes~
labels, etc., used in the remainder of the method. Generally, the cleavable site will be present in the moiety "Z" present within the R5 substituent in the formulae. Preferred cleavable sites arethoseidentifiedinU,S.PatentNo 5,118,605. Asexplainedinthatapplication,selectably cleavable sites include, for example, the ~;ullnwillg types of linkages:
O O
C ~ CH2 CH2 ~ (hydroxyla3nine-sensitive);
2û -C-NH- (b~se-sensitive);
O
-S- ( base-sens itive );
_5_5 _ ( thio1- s ens itive ); and Ol H Ol H
-CH--CH- ( periodate-sens itive ) .
N-hy h UAy ' ~ ' ~ (NHS) may be used to introduce the base-cleavable amide 30 bond into the reagent, while ethylene glycol bis(~ ~ ~ ~ 'yl succinate) may be used to create a hydroxyl~ , sen iitive linkage. bis[2-~ ' ~ yudl bùl~.~luAy)ethyl]sulfone (BSOCOES) may be used to create a ba~-sensitive sulfone linkage, ~ lyl tartarate 1 9 6 8 0 6 r, ~
~ WO 96/06104 P~ u / /6 (DST) may be used to introduce 1,2-diols cleavable by periodate, and dithiobis-~ 1~ ip, u~;~,.,atc) (DSP) may be used to provide thiol-cleavable disulfide bonds.
Methods of using these reagents to produce the desired cleavable linkage are well known and will be readily apparent to those skilled in the art of synthetic organic chemistry.
In the ar.,.c.. ,~.. ;u"cd ~ ~I.o~ , the moiety R9 represents a detectable label, such that cleavage of a linkage present within the spacer moiety Z will result in release of label .
Suitable labels which may be present at the R9 position in such a case include, for example, iu~ 4 ~, fluorescers, ' ' Imi~ srers~ dyes, enzymes, enzyme substrates, enzyme cofactors, enzyme inhibitors, enzyme subunits, metal ions, and the like. Illustrative specific 10 labels include fluorescein, rhodamine, Texas red, phycoerythrin, u".l,.llif~ , luminol, NADPH, a,b-~llr~iu~id~ .c, horseradish peroxidase, and the like.
Polynucleotide reagents useful as probes in hybridization assays may also be prepared by using the monomenc reagent (I) as a "branch poinl." In this way, probes containing branch points having the structural formula (V) - Ho r ~ Ai ~- ~ ~ P--~ ~ , ~, ,~ A ~ A ~ l-- G l~
O
1 ,5, ~
~r~--P--~-- ~ ~~11 o (V) wo96/06104 '1,~9 20- r~ lL ~/6 may be prepared, wherein DNA" DNA2, DNA3, R3, R4 and Rs are as defined above. Such 5 probes may be used, for example, in the ~ assays described in commoniy assigned U.S. Patent No. 5,124,246 to Urdea et al., entitied "Nucleic Acid Multimers and Amplified Nucleic Acid IIyblkl;~d~ivll Assays Using Same," PCT Publication Nos. W089/03891, and WO 92/02526. The latter application describes the comb-type branched multimers which are preferred in conjunction with the present method, and which are composed of a linear backbone and pendant sidechains; the backbone includes a segment that provides a specific hyl)li il~d~;UII site for anaiyte nucleic acid or nucleic acid bound to the analyte, whereas the pendant sidechains inciude iterations of a segment that provide specific Lrul;J;LdL;ull sites for a labeled probe.
In still another ~i.,Lo ihll~lL of the invention, the "abasic," or modified, site provided by monomeric reagent (I) may be used to enable synthesis of a ~ol~ .L.vliJu on a solid support. In this case, the reagent is bound to a solid support through the iinker arm at the I
position, i e., R9 represents a soiid support. As noted above, the linicage to the solid support may aiso be at the 3 position, at Rl Examples of solid supports include silica, Porasil C, polystyrene, controlled pore glass (CPG), kieselguhr, POIY(I~ GWYIG~Uid~
POIY(G~ llllUll ' ' ' ), polystyrene grafted onto pul.~ Lldlluulv~,Jl~ ;), cellulose, Sephadex LH-20 and Fractûsil 500. Nucleûtidic mûnomers are then added using standard DNA synthesis chemistry at the 3' and 5' positions. In some cases, i.e., to produce support-bound labeiied probes, it may be desirable to replace some nucleotidic monomers with labeiied monomers, e.g., the NJ-labelled cytidine derivatives described in commonly assigned U.S. Patent No. 5,093,232 to Urdea et al., entitled "Nucleic Acid Probes." Such monomers have the structurai formula (Vl) 2Ig680~ ' ~ WO96/06104 -21- r~ 1UI/6 R'~
R
(Vl) (~
1~
1 0 o~ ~ ~
~1~~o ~
OR
wherein R~ and Rl are as defined above;
R" is an optional linking moiety which, if present, contains an an~ide, thioether or disulfide linkage or a ~ .s,~ ;.." thereof;
2 o Rl2 is a reactive group d~,.iv~.Li~l,l,, with a detectable label, e.g., -NH~, -COOH or -SH;
R'3 is hydrogen, methyl, fluoro, bromo or iodo; and R'~ is either hydrogen, hydroxyl or protected hydroxyl.
In still another F~ .O- I : of the invention, I~UI ~ul,h,v~id~,3 are synthesized in which 25 themonomericreagent(I)maybeusedtochangethedirectionofsynthesis~e~g~from3~ 15'to 5' ~3' or vice versa. This is O ~ l by adding monomeric reagent (I) to the terminusofagrowingv~ vl;~lFchain~cappingeitherthe3~ors~terminalhydroxyl group with a capping group, typicaily an acyl capping group, and then using the l linker arm to continue synthesis in the reverse direction. Oligomers in which adjacent monomer units 3 0 are linked 3'-3' can also be prepared using reagent (I), by binding the ~ ;. IF to a solid support at R9, growing a single oligomer at the 5' position, capping exposed the WO 96106104 ~ 22 ~ J.,,J~ 6 group at the 5' terminus, and then growing a second oligomer at the 3' position. Such structures are illustrated in formulae (Il) and (III).
Synthetic Methods:
Scheme I illustrates the preferred method of ~yllLi. ,~;Lhlg monomeric reagents having the structural formula (1):
~ WO96/061~4 21968D6 p "~ o Scheme I
10 ~7~
o ' ~ ~
0~
9.' 1' ~ -~ ~
~r O7~ ~ v o ~ ¢7\~
o 1~ 7 S ,~ ~ 0 o ~5 , 30 ~s ~ X .1 ~!
o~
WO 96/06104 ?,~ 6 ~ / L l l6 In Scheme 1, 2-deoxy-D-, ;~ is used as the starting material. The three hydroxyl groups of the molecule are protected using an "R-CI" reagent or some other reagent suitable to protect free hydroxyl groups (e.g., benzoyl chloride or acetic anhydride) to provide 3 -OR groups at the I, 3 and 5 positions of the sugar. The product is isolated, and the l-OR group then replaced by reaction with a moiety (R9)~-X-~AH in the presence of an acid catalyst, followed by d.,~., uLeuL;on at the 3 and 5 positions using base. The S
position may then be selectively protected by reaction with Rl-CI, e.g., d;~ dlu~yLlilyl chloride, followed by reaction with a selected pl~ P at the 3 position to provide the desired phosphorus derivative.
The practice of the present invention will employ, unless otherwise indicated, ~,un~L;ull,~l techniques of synthetic organic chemistry, bio~,h~ L.y, molecular biology, and the like, which are within the skill of the art. Such techniques are explained fully in the literature. See, e.g., Sambrook, Fritsch & Maniatis, Molecular Clonin~ A J ol~nratnrv ~5~, Second Edition (1989); O8y~ ;de Svnthesis (M.J. Gait, ed., 1984), ~ç~ç~
Acid Ilvl ,. ;~ 1 (B.D. ~amcs & S.J. Higgins, eds.~ 1984); and a series, Methods in EI~YIIIOIUYY (Academic Press, Inc:). All patents, patent ~ , and I ' ' mentioned herein, both supra and i~u'ra, are hereby i~ ul~Jul~Ltd by reference.
2 0 It is to be understood that while the invention has been described in conjunction with the preferred specific ' ' thereof, that the description above as well as the exarnple which follows are intended to illustrate arLd not limit the scope of the invention. Other aspects, advantages and ,..n.l;i~ within the scope ofthe invention will be apparent to those skilled in the art to which the invention pertains.
In the following example, efforts have been made to insure accuracy with respect to numbers used (e.g., amounts, Ltl..~ LU.l:, etc.) but some c l~ ; ..;~1 error and deviation should be accounted for. Unless indicated otherwise, LtllllJ.,l~Lul~ is in degrees C and pressure is at or near ,a~ 8 2l968~6 .' ~',, ' ' ~ P~ 1 16 Exam~le I
1,3,5-Tris-O-acyl-2-deoxy-D-,;i,uru,~,lu~c was readily synthesized by treating CU~ILI.~ Y available 2-deoxy-D-. ;1,. ,~: " .., ~. .~ with a large excess of acetic anhydride or benzoyl chloride in pyridine. Both 1,3,5-O-trisacetyi- and trisbenzoyl-2-deoxy-5 D-,;i,uru"..oaG couid be ~ G~l y ' "' ~ firom ethanol. The acetyl derivative was mainiy the aipha-isomer, and the benzoyl derivative gave the two isomers in 1/1 ratio. No pyrancside derivative was formed ( less than 5% ).
1,3,5-O-tris(TBDMS)-2'-deoxy-D-, ;1 ", ~ 5itlr- was synthesized from ieU~Lyl ;iJo:~G by reaction with t-buLyl ihl.~..llybil~l ~,1 iu,;dc';"..d, ~JIc/DMF.
The anomeric acetai was readily exchanged with an alcohol in the presence of an acid catalyst, such as ZnBr2, to give the alcohol derivative of either 3,5-0-diacyl- or 3,5-O- di-TBDMS -2'-deoxy-, ;1 " ,1; ", ~f Removal of the 3,5-O-protecting groups with base (methanoi/lM K2CO3 for acyi) or fluoride ions (IM LGLI.LI,UIYI~"U,-U....I-., fluoride in THF
for TBDMS) gave the substituted 2'-deoxy- ~;burul~ uac derivatives.
Aicoholscontainingvarious li, li.~ ; havebeen;.. ~,u,pu,dltdthisway.
R~,yl~"c~t~L;vG examples are 4-1ll~,.~.ylu~y-".-lJu--yllJ.,.~yl, 4-1ut~uyll~.l..,Lllyl~
TFA-NH-alicyl(aryl), and N-(4~u~ ub~ Iu~y~;~uul~yl)/
~;~IOC-6-aminohexyl. They were aii prepared directly from the CUIlGyUlld;ll~; alcohol and 1,3,5-tri-0-acyl-2'-deoxy-D-,ii,uru..l,,uaG.
2 0 Preparation of S-trityl- I l-mercapto- I-undecyl was achieved via the 11 -bromo-l-undecyl derivative: After preparation of the 11 -bromo- I -undecyl 3,5-di-O-acetyl-2-deoxy-D-,ii,uL~l~u,uaG reaction with ~iLyh~,.l , (Tr-SH) in the presence of base (one equivaient of aq. NaOH ) afforded S-trityl- 11-u~ yLuulld~,~.yl 2-deoxy-D-.;l,~ r~ Aiternatively, S-Tr-ll-mercapto-l-undecanol could be prepared and used as the aicohol component. Aiternatively, it is possible to incorporate aicohols containing a disulfite, -S-S-. The O-levuiinyl-l l-oxo-undecyl derivative was prepared via the 11 -bromo derivative. After removai of the acetyl ~roups, of bromine with the Cs-salt of levulinic acid afforded O-levulinyl-l l -oxy-undecyl 2-deoxy-D-,ii,, ~ ~ Aiternatively, preformed O-levulinyl-l l-oxy-l-undecanol 3 0 could be used as the alcohol component.
The appropriate alicyl 2-deoxy-D-~ il,uru- a"uaidG analogs were converted to theS-DMT derivatives using stamdard literature procedures. The two anomeric aklGu;Su..._l a ,, . .. . ...... ... . _ WO 96/06104 ?~ 26 P~ 16 gave rise to DMT species with quite different mobilities during silica gel clll~ " . ', .
All DMT ' were purified by silica gel ~,LI ulllaLu~ lly, and the two anomeric ~Lcl CU;~UIII~ were readily separated. The various DMT ;"~ were converted to the 3-0-N,N-d;;~u~luuyl~.yGIlu~,;llyl-~ lln~ using standard literature procedures, and they could be used like normal nucleoside uy.-llu~ yl~ during automated synthesis.
Removal of protecting groups from chemically synthesized 1;". ~ ul; '~ ~ required only minimal changes to the standard procedures.
~ ..,.,,h~ !u~y~,albu~l ~ lu~l~yl 2-deoxy-D-~ u~ Hydrolysis of methyl ester and succinate linkage to support was carried out with water/TEA/dioxane (I :1:10 v/v; 13 hours) prior to exposure to ammonium hydroxide.
TFA/FMOC-NH-alkyl required only standard deprotection with ammonium hydroxide. For 4-1l;LIu~L~ ;llyl and N-(4--~ ub.,.l~lu~y-carbonyl)-6-~lli-lùli~yl.
reduction of the nitro group to an anilino group was conducted with 0.1 M sodium15 dithionitellM TEAB/ dioxane for S hours, washed, and then deprotected with ammonium hydroxide to give the free anilino- and amino derivatized oligomer, l~ ,.,Li~ , which was purified by PAGE.
On support treatment with HPAA reagent, the DNA synthesis can be continued on the same support to produce branched oligomers. With proper choice of side-arm length, 2 0 the monomer is usefiul for making 3'-3' linked oli~,uu~ uLid~ for cross-over triple helix formation. An exarnple is O-levulinyl-2-oxyethyl S-DMT-0-2-deoxy-D-, il ,. .ll .. ~
3'-O-succ-CPG; the first strand is synthesized using 5'-DMT, capped, the levulinyl group removed and synthesis continued at the 2-h~d~u~ ;h~l side-chain. Deprotection gives the desired 5'-DNAI-3'3'-DNA2-5' oligomer.
Claims (11)
1. A reagent having the structural formula wherein:
R1 is selected from the group consisting of hydrogen, acid-sensitive, base-stable protecting groups and acyl capping groups;
R2 is a phosphorus derivative selected to enable addition of the reagent to a molecular species containing a free hydroxyl group, or is a linkage to a solid support;
R3 is selected from the group consisting of hydrogen, hydroxyl, sulfhydryl, halogeno, amino, alkyl, allyl, -OR6 wherein R6 is alkyl, allyl, silyl or phosphate;
R4 is either hydrogen or -(CH2)mOR7 wherein R7 is alkyl or -(CO)R8, R8 is alkyl, and m is an integer in the range of 0 to 12 inclusive;
R5 is -A-Z-X(R9)n;
A is oxygen, sulfur or methylene;
Z is arylene, C6-C18 aralkylene or C1-C12 alkylene containing 0 to 6 heteroatomsselected from the group consisting of O, S, N, Si and Se and 0 to 6 linkages selected from the group consisting of -CO-, -COO-, -CONH-, -NHCO-, -S-S-, -SO2-, -CH(OH)-CH(OH)-, -CH(OR4)-CH(OR4)-, -O-PO(O)-O-, -O-PO(R4)-, -O-PO(OR4)-O-, -O-PO(OR4)-R5- and -PO(OR4)-O-R5- in which R4 is lower alkyl and R5 is lower alkylene, and, if Z is aralkylene or alkylene, containing 0 to 3 unsaturated bonds;
X is selected from the group consisting of -NH-, -CONH-, -NHCO-, -CO-, -S- and -Si~;
R9 is hydrogen, a protecting group, a detectable label, or, unless X is -Si~, a solid support; and n is 1 when X is -NH-, -CONH-, -NHCO-, -CO-, or -S-, and is 3 when X is -Si~, with the proviso that if R2 represents a linkage to a solid support, R9 is hydrogen, a protecting group, or a detectable label.
R1 is selected from the group consisting of hydrogen, acid-sensitive, base-stable protecting groups and acyl capping groups;
R2 is a phosphorus derivative selected to enable addition of the reagent to a molecular species containing a free hydroxyl group, or is a linkage to a solid support;
R3 is selected from the group consisting of hydrogen, hydroxyl, sulfhydryl, halogeno, amino, alkyl, allyl, -OR6 wherein R6 is alkyl, allyl, silyl or phosphate;
R4 is either hydrogen or -(CH2)mOR7 wherein R7 is alkyl or -(CO)R8, R8 is alkyl, and m is an integer in the range of 0 to 12 inclusive;
R5 is -A-Z-X(R9)n;
A is oxygen, sulfur or methylene;
Z is arylene, C6-C18 aralkylene or C1-C12 alkylene containing 0 to 6 heteroatomsselected from the group consisting of O, S, N, Si and Se and 0 to 6 linkages selected from the group consisting of -CO-, -COO-, -CONH-, -NHCO-, -S-S-, -SO2-, -CH(OH)-CH(OH)-, -CH(OR4)-CH(OR4)-, -O-PO(O)-O-, -O-PO(R4)-, -O-PO(OR4)-O-, -O-PO(OR4)-R5- and -PO(OR4)-O-R5- in which R4 is lower alkyl and R5 is lower alkylene, and, if Z is aralkylene or alkylene, containing 0 to 3 unsaturated bonds;
X is selected from the group consisting of -NH-, -CONH-, -NHCO-, -CO-, -S- and -Si~;
R9 is hydrogen, a protecting group, a detectable label, or, unless X is -Si~, a solid support; and n is 1 when X is -NH-, -CONH-, -NHCO-, -CO-, or -S-, and is 3 when X is -Si~, with the proviso that if R2 represents a linkage to a solid support, R9 is hydrogen, a protecting group, or a detectable label.
2. The reagent of claim 1, wherein A is oxygen.
3. The reagent of claim 2, wherein R9 is trityl.
4. The reagent of claim 2, wherein R9 is a label.
5. A reagent having the structural formula wherein:
R1 is selected from the group consisting of hydrogen and acid-sensitive, base-stable protecting groups;
R2 is is selected from the group consisting of phosphoramidites, phosphotriesters, phosphodiesters, phosphites, H-phosphonates and phosphorothioates;
A is oxygen, sulfur or methylene;
Z is a hydrocarbyl or oxyhydrocarbyl spacer moiety containing 1 to 18 carbon atoms and 0 to 6 oxygen atoms;
X is selected from the group consisting of -NH- and -S-; and R9 is a protecting group.
R1 is selected from the group consisting of hydrogen and acid-sensitive, base-stable protecting groups;
R2 is is selected from the group consisting of phosphoramidites, phosphotriesters, phosphodiesters, phosphites, H-phosphonates and phosphorothioates;
A is oxygen, sulfur or methylene;
Z is a hydrocarbyl or oxyhydrocarbyl spacer moiety containing 1 to 18 carbon atoms and 0 to 6 oxygen atoms;
X is selected from the group consisting of -NH- and -S-; and R9 is a protecting group.
6. A polynucleotide reagent having the structural formula wherein:
DNA1 is a first segment of DNA;
DNA2 is a second segment of DNA;
R3 is selected from the group consisting of hydrogen, hydroxyl, sulfhydryl, halogeno, amino, alkyl, allyl, OR6 wherein R6 is alkyl, allyl, silyl or phosphate;
R4 is either hydrogen or -(CH2)mOR7 wherein R7 is alkyl or -(CO)R8, R8 is alkyl, and m is an integer in the range of 0 to 12 inclusive;
R5 is -A-Z-X(R9)n;
A is oxygen, sulfur or methylene;
Z is arylene, C6-C18 aralkylene or C1-C12 alkylene containing 0 to heteroatoms selected from the group consisting of O, S, N,Si and Se and 0 to 6 linkages selected from the group consisting of-CO-, -COO-, -CONH-, -NHCO-, -S-S-, -SO2-, -CH(OH)-CH(OH)-, -CH(OR4)-CH(OR4)-, -O-PO(O)-O-, -O-PO(R4)-, -O-PO(OR4)-O-, -O-PO(OR4)-R5- and -PO(OR4)-O-R5- in which R4 is lower alkyl and R5 is lower alkylene, and, if Z is aralkylene or alkylene, containing 0 to 3 unsaturated bonds;
X is selected from the group consisting of -NH-, -CONH-, -NHCO-, -CO-, -S- and -Si~
R9 is hydrogen, a protecting group, or a detectable label; and n is 1 when X is -NH-, -CONH-, -NHCO-, -CO-, or -S-, and is 3 when X is -Si~.
DNA1 is a first segment of DNA;
DNA2 is a second segment of DNA;
R3 is selected from the group consisting of hydrogen, hydroxyl, sulfhydryl, halogeno, amino, alkyl, allyl, OR6 wherein R6 is alkyl, allyl, silyl or phosphate;
R4 is either hydrogen or -(CH2)mOR7 wherein R7 is alkyl or -(CO)R8, R8 is alkyl, and m is an integer in the range of 0 to 12 inclusive;
R5 is -A-Z-X(R9)n;
A is oxygen, sulfur or methylene;
Z is arylene, C6-C18 aralkylene or C1-C12 alkylene containing 0 to heteroatoms selected from the group consisting of O, S, N,Si and Se and 0 to 6 linkages selected from the group consisting of-CO-, -COO-, -CONH-, -NHCO-, -S-S-, -SO2-, -CH(OH)-CH(OH)-, -CH(OR4)-CH(OR4)-, -O-PO(O)-O-, -O-PO(R4)-, -O-PO(OR4)-O-, -O-PO(OR4)-R5- and -PO(OR4)-O-R5- in which R4 is lower alkyl and R5 is lower alkylene, and, if Z is aralkylene or alkylene, containing 0 to 3 unsaturated bonds;
X is selected from the group consisting of -NH-, -CONH-, -NHCO-, -CO-, -S- and -Si~
R9 is hydrogen, a protecting group, or a detectable label; and n is 1 when X is -NH-, -CONH-, -NHCO-, -CO-, or -S-, and is 3 when X is -Si~.
7. A polynucleotide reagent having the structural formula wherein DNA1 is a first segment of DNA;
DNA2 is a second segment of DNA;
R3 is selected from the group consisting of hydrogen, hydroxyl, sulfhydryl, halogeno, amino, alkyl, allyl, OR6 wherein R6 is alkyl, allyl, silyl or phosphate;
R4 is either hydrogen or -(CH2)mOR7 wherein R7 is alkyl or -(CO)R8, R8 is alkyl, and m is an integer in the range of 0 to 12 inclusive, A is oxygen, sulfur or methylene; and Z is arylene, C6-C18 aralkylene or C1-C12 alkylene containing 0 to 6 heteroatomsselected from the group consisting of O, S, N, Si and Se and 0 to 6 linkages selected from the group consisting of -CO-,-COO-, -CONH-, -NHCO-, -S-S-, -SO2, -CH(OH)-CH(OH)-, -CH(OR4)-CH(OR4)-, -O-PO(O-)-O-, -O-PO(R4)-, -O-PO(OR4)-O-, -O-PO(OR4)-R5- and -PO(OR4)-O-R5- in which R4 is lower alkyl and R5 is lower alkylene, and, if Z is aralkylene or alkalene, containing 0 to 3 unsatursted bonds.
DNA2 is a second segment of DNA;
R3 is selected from the group consisting of hydrogen, hydroxyl, sulfhydryl, halogeno, amino, alkyl, allyl, OR6 wherein R6 is alkyl, allyl, silyl or phosphate;
R4 is either hydrogen or -(CH2)mOR7 wherein R7 is alkyl or -(CO)R8, R8 is alkyl, and m is an integer in the range of 0 to 12 inclusive, A is oxygen, sulfur or methylene; and Z is arylene, C6-C18 aralkylene or C1-C12 alkylene containing 0 to 6 heteroatomsselected from the group consisting of O, S, N, Si and Se and 0 to 6 linkages selected from the group consisting of -CO-,-COO-, -CONH-, -NHCO-, -S-S-, -SO2, -CH(OH)-CH(OH)-, -CH(OR4)-CH(OR4)-, -O-PO(O-)-O-, -O-PO(R4)-, -O-PO(OR4)-O-, -O-PO(OR4)-R5- and -PO(OR4)-O-R5- in which R4 is lower alkyl and R5 is lower alkylene, and, if Z is aralkylene or alkalene, containing 0 to 3 unsatursted bonds.
8. A polynucleotide reagent having the structural formula wherein DNA1 is a first segment of DNA;
DNA2 is a second segment of DNA;
R3 is selected from the group consisting of hydrogen, hydroxyl, sulfhydryl, halogeno, amino, alkyl, allyl, -OR6 wherein R6 is alkyl, allyl, silyl or phosphate;
R4 is either hydrogen or -(CH2)mOR7 wherein R7 is alkyl or -(CO)R3, R8 is alkyl, and m is an integer in the range of 0 to 12 inclusive, A is oxygen, sulfur or methylene; and Z is arylene, C6-C18 aralkylene or C1-C12 alkylene containing 0 to 6 heteroatomsselected from the group consisting of O, S, N, Si and Se and 0 to 6 linkages selected from the group consisting of-CO-,-COO-, -CONH-, -NHCO-, -S-S-, -SO2-, -CH(OH)-CH(OH)-, -CH(OR4)-CH(OR4)-, -O-PO(O)-O-, -O-PO(R4)-,-O-PO(OR4)-O-, -O-PO(OR4)-R5- and -PO(OR4)-O-R5- in which R4 is lower alkyl and R5 is lower alkylene, and, if Z is araikylene or alkylene, containing 0 to 3 unsaturated bonds.
DNA2 is a second segment of DNA;
R3 is selected from the group consisting of hydrogen, hydroxyl, sulfhydryl, halogeno, amino, alkyl, allyl, -OR6 wherein R6 is alkyl, allyl, silyl or phosphate;
R4 is either hydrogen or -(CH2)mOR7 wherein R7 is alkyl or -(CO)R3, R8 is alkyl, and m is an integer in the range of 0 to 12 inclusive, A is oxygen, sulfur or methylene; and Z is arylene, C6-C18 aralkylene or C1-C12 alkylene containing 0 to 6 heteroatomsselected from the group consisting of O, S, N, Si and Se and 0 to 6 linkages selected from the group consisting of-CO-,-COO-, -CONH-, -NHCO-, -S-S-, -SO2-, -CH(OH)-CH(OH)-, -CH(OR4)-CH(OR4)-, -O-PO(O)-O-, -O-PO(R4)-,-O-PO(OR4)-O-, -O-PO(OR4)-R5- and -PO(OR4)-O-R5- in which R4 is lower alkyl and R5 is lower alkylene, and, if Z is araikylene or alkylene, containing 0 to 3 unsaturated bonds.
9. A branched polynucleotide reagent having the structural formula wherein:
DNA1 is a first segment of DNA;
DNA2 is a second segment of DNA;
DNA3 is a third segment of DNA;
R3 is selected from the group consisting of hydrogen, hydroxyl, sulfhydryl, halogeno, amino, alkyl, allyl, -OR6 wherein R6 is alkyl, allyl, silyl or phosphate;
R4 is either hydrogen or -(CH2)mOR7 wherein R7 is alkyl or -(CO)R8, R8 is alkyl, and m is an integer in the range of 0 to 12 inclusive;
A is oxygen, sulfur or methylene; and Z is arylene, C6-C18 aralkylene or C1-C12 alkylene containing 0 to 6 heteroatomsselected from the group consisting of O, S, N, Si and Se and 0 to 6 linkages selected from the group consisting of -CO-, -COO-, -CONH-, -NHCO-, -S-S-, -SO2-, -CH(OH)-CH(OH)-, -CH(OR4)-CH(OR4)-, -O-PO(O-)-O-, -O-PO(R4)-, -O-PO(OR4)-O-, -O-PO(OR4)-R5- and -PO(OR4)-O-R5- in which R4 is lower alkyl and R5 is lower alkylene, and, if Z is aralkylene or alkylene, containing 0 to 3 unsaturated bonds.
DNA1 is a first segment of DNA;
DNA2 is a second segment of DNA;
DNA3 is a third segment of DNA;
R3 is selected from the group consisting of hydrogen, hydroxyl, sulfhydryl, halogeno, amino, alkyl, allyl, -OR6 wherein R6 is alkyl, allyl, silyl or phosphate;
R4 is either hydrogen or -(CH2)mOR7 wherein R7 is alkyl or -(CO)R8, R8 is alkyl, and m is an integer in the range of 0 to 12 inclusive;
A is oxygen, sulfur or methylene; and Z is arylene, C6-C18 aralkylene or C1-C12 alkylene containing 0 to 6 heteroatomsselected from the group consisting of O, S, N, Si and Se and 0 to 6 linkages selected from the group consisting of -CO-, -COO-, -CONH-, -NHCO-, -S-S-, -SO2-, -CH(OH)-CH(OH)-, -CH(OR4)-CH(OR4)-, -O-PO(O-)-O-, -O-PO(R4)-, -O-PO(OR4)-O-, -O-PO(OR4)-R5- and -PO(OR4)-O-R5- in which R4 is lower alkyl and R5 is lower alkylene, and, if Z is aralkylene or alkylene, containing 0 to 3 unsaturated bonds.
10. In a method for making a polynucleotide reagent comprising sequentially coupling nucleotidic monomers to a growing oligonucleotide chain, the improvement which comprises introducing an abasic site into the polynucleotide reagent by replacing a fraction of the nucleotidic monomers with the reagent of claim 9.
11. A method for making branched DNA, comprising: (a) sequentially coupling nucleotidic monomers to a growing oligonucleotide chain; (b) introducing branch points into the chain during step (a) by replacing a fraction of the nucleotidic monomers with monomeric reagents having linker arms at the 1 position; (c) sequentially adding nucleotidic monomers to the termini of the linker arms, wherein each of the monomeric reagents used in step (b) comprises a reagent of claim 1.
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US08/296,368 | 1994-08-25 | ||
US08/296,368 US5597909A (en) | 1994-08-25 | 1994-08-25 | Polynucleotide reagents containing modified deoxyribose moieties, and associated methods of synthesis and use |
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EP (1) | EP0777674B1 (en) |
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CA (1) | CA2196806A1 (en) |
DE (1) | DE69524232T2 (en) |
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Families Citing this family (747)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6335434B1 (en) | 1998-06-16 | 2002-01-01 | Isis Pharmaceuticals, Inc., | Nucleosidic and non-nucleosidic folate conjugates |
US8153602B1 (en) | 1991-11-19 | 2012-04-10 | Isis Pharmaceuticals, Inc. | Composition and methods for the pulmonary delivery of nucleic acids |
JP3484197B2 (en) | 1993-09-03 | 2004-01-06 | アイシス・ファーマシューティカルス・インコーポレーテッド | Amine derivatized nucleosides and oligonucleosides |
JP2000500740A (en) * | 1995-10-19 | 2000-01-25 | プロリゴ・エルエルシー | Solution-phase synthesis of oligonucleotides |
US9096636B2 (en) | 1996-06-06 | 2015-08-04 | Isis Pharmaceuticals, Inc. | Chimeric oligomeric compounds and their use in gene modulation |
US5898031A (en) * | 1996-06-06 | 1999-04-27 | Isis Pharmaceuticals, Inc. | Oligoribonucleotides for cleaving RNA |
US20030044941A1 (en) | 1996-06-06 | 2003-03-06 | Crooke Stanley T. | Human RNase III and compositions and uses thereof |
US20040147022A1 (en) * | 1996-06-06 | 2004-07-29 | Baker Brenda F. | 2'-methoxy substituted oligomeric compounds and compositions for use in gene modulations |
WO2005121368A1 (en) * | 2004-06-03 | 2005-12-22 | Isis Pharmaceuticals, Inc. | Chimeric gapped oligomeric compositions |
US7812149B2 (en) * | 1996-06-06 | 2010-10-12 | Isis Pharmaceuticals, Inc. | 2′-Fluoro substituted oligomeric compounds and compositions for use in gene modulations |
US20040203024A1 (en) * | 1996-06-06 | 2004-10-14 | Baker Brenda F. | Modified oligonucleotides for use in RNA interference |
US20050053976A1 (en) * | 1996-06-06 | 2005-03-10 | Baker Brenda F. | Chimeric oligomeric compounds and their use in gene modulation |
US5853993A (en) * | 1996-10-21 | 1998-12-29 | Hewlett-Packard Company | Signal enhancement method and kit |
US6187536B1 (en) | 1997-02-18 | 2001-02-13 | Thomas Jefferson University | Methods of identifying and detecting pancreatic cancer |
IL132377A0 (en) * | 1997-04-21 | 2001-03-19 | Proligo Llc | Method for solution phase synthesis of oligonucleotides |
JP2002510319A (en) | 1997-07-01 | 2002-04-02 | アイシス・ファーマシューティカルス・インコーポレーテッド | Compositions and methods for delivery of oligonucleotides through the gastrointestinal tract |
US7135333B1 (en) | 1997-08-07 | 2006-11-14 | Thomas Jefferson University | Compositions that specifically bind to colorectal cancer cells and methods of using the same |
US6120995A (en) * | 1997-08-07 | 2000-09-19 | Thomas Jefferson University | Compositions that specifically bind to colorectal cancer cells and methods of using the same |
WO1999049082A2 (en) | 1998-03-23 | 1999-09-30 | Invitrogen Corporation | Modified nucleotides and methods useful for nucleic acid sequencing |
US20040186071A1 (en) * | 1998-04-13 | 2004-09-23 | Bennett C. Frank | Antisense modulation of CD40 expression |
US7321828B2 (en) * | 1998-04-13 | 2008-01-22 | Isis Pharmaceuticals, Inc. | System of components for preparing oligonucleotides |
WO1999060167A1 (en) * | 1998-05-21 | 1999-11-25 | Isis Pharmaceuticals, Inc. | Compositions and methods for topical delivery of oligonucleotides |
EP1469009A2 (en) * | 1998-05-21 | 2004-10-20 | Isis Parmaceuticals, Inc. | Compositions and methods for non-parenteral delivery of oligonucleotides |
US6335439B1 (en) | 1998-06-11 | 2002-01-01 | Isis Pharmaceuticals, Inc. | Method of preparing phosphoramidites |
US6225293B1 (en) | 1998-09-02 | 2001-05-01 | Isis Pharmaceuticals, Inc. | Methods and compounds for tracking the biodistribution of macromolecule-carrier combinations |
US6077709A (en) | 1998-09-29 | 2000-06-20 | Isis Pharmaceuticals Inc. | Antisense modulation of Survivin expression |
US6203989B1 (en) | 1998-09-30 | 2001-03-20 | Affymetrix, Inc. | Methods and compositions for amplifying detectable signals in specific binding assays |
US6300320B1 (en) | 1999-01-05 | 2001-10-09 | Isis Pharmaceuticals, Inc. | Modulation of c-jun using inhibitors of protein kinase C |
US7098192B2 (en) | 1999-04-08 | 2006-08-29 | Isis Pharmaceuticals, Inc. | Antisense oligonucleotide modulation of STAT3 expression |
US20060275782A1 (en) * | 1999-04-20 | 2006-12-07 | Illumina, Inc. | Detection of nucleic acid reactions on bead arrays |
EP1196630B2 (en) | 1999-04-20 | 2018-10-17 | Illumina, Inc. | Detection of nucleic acid reactions on bead arrays |
US20030207295A1 (en) * | 1999-04-20 | 2003-11-06 | Kevin Gunderson | Detection of nucleic acid reactions on bead arrays |
US8481268B2 (en) | 1999-05-21 | 2013-07-09 | Illumina, Inc. | Use of microfluidic systems in the detection of target analytes using microsphere arrays |
US8080380B2 (en) * | 1999-05-21 | 2011-12-20 | Illumina, Inc. | Use of microfluidic systems in the detection of target analytes using microsphere arrays |
US6656730B1 (en) | 1999-06-15 | 2003-12-02 | Isis Pharmaceuticals, Inc. | Oligonucleotides conjugated to protein-binding drugs |
US6147200A (en) * | 1999-08-19 | 2000-11-14 | Isis Pharmaceuticals, Inc. | 2'-O-acetamido modified monomers and oligomers |
US6617442B1 (en) | 1999-09-30 | 2003-09-09 | Isis Pharmaceuticals, Inc. | Human Rnase H1 and oligonucleotide compositions thereof |
US7332275B2 (en) | 1999-10-13 | 2008-02-19 | Sequenom, Inc. | Methods for detecting methylated nucleotides |
US6261840B1 (en) | 2000-01-18 | 2001-07-17 | Isis Pharmaceuticals, Inc. | Antisense modulation of PTP1B expression |
US20020055479A1 (en) | 2000-01-18 | 2002-05-09 | Cowsert Lex M. | Antisense modulation of PTP1B expression |
US20030176385A1 (en) * | 2000-02-15 | 2003-09-18 | Jingfang Ju | Antisense modulation of protein expression |
ES2548381T3 (en) | 2000-03-27 | 2015-10-16 | Thomas Jefferson University | Compositions for the treatment and imaging of stomach and esophageal cancer cells |
AU2001253408C1 (en) * | 2000-04-13 | 2008-12-04 | Mervyn Merrilees | Therapeutic compounds and methods for formulating V3, A Versican Isoform |
US6680172B1 (en) | 2000-05-16 | 2004-01-20 | Regents Of The University Of Michigan | Treatments and markers for cancers of the central nervous system |
US20060166227A1 (en) * | 2000-06-20 | 2006-07-27 | Stephen Kingsmore | Protein expression profiling |
US6323009B1 (en) * | 2000-06-28 | 2001-11-27 | Molecular Staging, Inc. | Multiply-primed amplification of nucleic acid sequences |
US6958214B2 (en) | 2000-07-10 | 2005-10-25 | Sequenom, Inc. | Polymorphic kinase anchor proteins and nucleic acids encoding the same |
US8568766B2 (en) * | 2000-08-24 | 2013-10-29 | Gattadahalli M. Anantharamaiah | Peptides and peptide mimetics to treat pathologies associated with eye disease |
AU2001292728A1 (en) * | 2000-09-18 | 2002-03-26 | Thomas Jefferson University | Compositions and methods for identifying and targeting stomach and esophageal cancer cells |
US20020115058A1 (en) * | 2000-09-22 | 2002-08-22 | Pedersen Finn Skou | Methods for diagnosis and treatment of diseases associated with altered expression of Pik3r1 |
US20030044803A1 (en) * | 2000-09-22 | 2003-03-06 | Pedersen Finn Skou | Methods for diagnosis and treatment of diseases associated with altered expression of JAK1 |
US20020164576A1 (en) * | 2000-09-22 | 2002-11-07 | Pedersen Finn Skou | Methods for diagnosis and treatment of diseases associated with altered expression of Nrf2 |
JP5305553B2 (en) | 2000-10-12 | 2013-10-02 | ユニバーシティー オブ ロチェスター | Composition for inhibiting the growth of cancer cells |
US7645441B2 (en) | 2000-12-22 | 2010-01-12 | Sagres Discovery Inc. | Compositions and methods in cancer associated with altered expression of PRLR |
US20030165878A1 (en) * | 2000-12-22 | 2003-09-04 | Morris David W. | Novel compositions and methods in cancer associated with altered expression of MCM3AP |
US7892730B2 (en) * | 2000-12-22 | 2011-02-22 | Sagres Discovery, Inc. | Compositions and methods for cancer |
US20030232334A1 (en) | 2000-12-22 | 2003-12-18 | Morris David W. | Novel compositions and methods for cancer |
US20030087252A1 (en) * | 2000-12-22 | 2003-05-08 | Morris David W. | Novel compositions and methods in cancer associated with altered expression of PRDM11 |
US7700274B2 (en) * | 2000-12-22 | 2010-04-20 | Sagres Discovery, Inc. | Compositions and methods in cancer associated with altered expression of KCNJ9 |
US7820447B2 (en) | 2000-12-22 | 2010-10-26 | Sagres Discovery Inc. | Compositions and methods for cancer |
US20030099963A1 (en) * | 2000-12-22 | 2003-05-29 | Morris David W. | Novel compositions and methods in cancer associated with altered expression of TBX21 |
US7767802B2 (en) | 2001-01-09 | 2010-08-03 | Alnylam Pharmaceuticals, Inc. | Compositions and methods for inhibiting expression of anti-apoptotic genes |
US6573051B2 (en) * | 2001-03-09 | 2003-06-03 | Molecular Staging, Inc. | Open circle probes with intramolecular stem structures |
US20030092157A1 (en) * | 2001-03-16 | 2003-05-15 | Hayden Michael R. | Compositions, screening systems and methods for modulating HDL cholesterol and triglyceride levels |
US6677120B2 (en) * | 2001-03-30 | 2004-01-13 | Isis Pharmaceuticals, Inc. | Building blocks for the solution phase synthesis of oligonucleotides |
US20030191073A1 (en) | 2001-11-07 | 2003-10-09 | Challita-Eid Pia M. | Nucleic acid and corresponding protein entitled 161P2F10B useful in treatment and detection of cancer |
WO2002097123A2 (en) * | 2001-05-25 | 2002-12-05 | Xenon Genetics, Inc. | Diagnostic methods for cardiovascular disease, low hdl-cholesterol levels, and high triglyceride levels |
US7803915B2 (en) * | 2001-06-20 | 2010-09-28 | Genentech, Inc. | Antibody compositions for the diagnosis and treatment of tumor |
CA2633171C (en) | 2001-06-20 | 2012-11-20 | Genentech, Inc. | Antibodies against tumor-associated antigenic target (tat) polypeptides |
US20050107595A1 (en) * | 2001-06-20 | 2005-05-19 | Genentech, Inc. | Compositions and methods for the diagnosis and treatment of tumor |
CA2790034A1 (en) | 2001-06-21 | 2003-01-03 | Isis Pharmaceuticals, Inc. | Antisense modulation of superoxide dismutase 1, soluble expression |
US7425545B2 (en) | 2001-07-25 | 2008-09-16 | Isis Pharmaceuticals, Inc. | Modulation of C-reactive protein expression |
US6964950B2 (en) | 2001-07-25 | 2005-11-15 | Isis Pharmaceuticals, Inc. | Antisense modulation of C-reactive protein expression |
US20030096772A1 (en) | 2001-07-30 | 2003-05-22 | Crooke Rosanne M. | Antisense modulation of acyl CoA cholesterol acyltransferase-2 expression |
US7407943B2 (en) | 2001-08-01 | 2008-08-05 | Isis Pharmaceuticals, Inc. | Antisense modulation of apolipoprotein B expression |
US7227014B2 (en) | 2001-08-07 | 2007-06-05 | Isis Pharmaceuticals, Inc. | Antisense modulation of apolipoprotein (a) expression |
NZ573831A (en) | 2001-09-18 | 2010-07-30 | Genentech Inc | Compositions and methods for the diagnosis and treatment of tumor, particularly breast tumor - TAT193 |
US20070098728A1 (en) * | 2001-09-24 | 2007-05-03 | Pedersen Finn S | Novel compositions and methods in cancer |
NZ566396A (en) | 2001-10-09 | 2009-07-31 | Isis Pharmaceuticals Inc | Antisense modulation of insulin-like growth factor binding protein 5 expressions |
US6750019B2 (en) | 2001-10-09 | 2004-06-15 | Isis Pharmaceuticals, Inc. | Antisense modulation of insulin-like growth factor binding protein 5 expression |
US20040166490A1 (en) * | 2002-12-17 | 2004-08-26 | Morris David W. | Novel therapeutic targets in cancer |
US20040126762A1 (en) * | 2002-12-17 | 2004-07-01 | Morris David W. | Novel compositions and methods in cancer |
AU2002364945A1 (en) * | 2001-10-25 | 2003-07-09 | Neurogenetics, Inc. | Genes and polymorphisms on chromosome 10 associated with alzheimer's disease and other neurodegenerative diseases |
US20030224380A1 (en) * | 2001-10-25 | 2003-12-04 | The General Hospital Corporation | Genes and polymorphisms on chromosome 10 associated with Alzheimer's disease and other neurodegenerative diseases |
US20030170678A1 (en) * | 2001-10-25 | 2003-09-11 | Neurogenetics, Inc. | Genetic markers for Alzheimer's disease and methods using the same |
US20040197778A1 (en) * | 2002-12-26 | 2004-10-07 | Sagres Discovery, Inc. | Novel compositions and methods in cancer |
US20040180344A1 (en) * | 2003-03-14 | 2004-09-16 | Morris David W. | Novel therapeutic targets in cancer |
US20060040262A1 (en) * | 2002-12-27 | 2006-02-23 | Morris David W | Novel compositions and methods in cancer |
US6965025B2 (en) | 2001-12-10 | 2005-11-15 | Isis Pharmaceuticals, Inc. | Antisense modulation of connective tissue growth factor expression |
NZ533933A (en) | 2002-01-02 | 2008-06-30 | Genentech Inc | Compositions and methods for the diagnosis and treatment of glioma tumor |
US7553619B2 (en) * | 2002-02-08 | 2009-06-30 | Qiagen Gmbh | Detection method using dissociated rolling circle amplification |
US20040023267A1 (en) * | 2002-03-21 | 2004-02-05 | Morris David W. | Novel compositions and methods in cancer |
US7169916B2 (en) * | 2002-04-01 | 2007-01-30 | Isis Pharmaceuticals, Inc. | Chloral-free DCA in oligonucleotide synthesis |
CA2481507A1 (en) | 2002-04-16 | 2003-10-30 | Genentech, Inc. | Compositions and methods for the diagnosis and treatment of tumor |
CA2484676A1 (en) * | 2002-05-03 | 2003-11-13 | Sequenom, Inc. | Kinase anchor protein muteins, peptides thereof, and related methods |
US7176181B2 (en) * | 2002-05-21 | 2007-02-13 | Yeda Research And Development Co. Ltd. | Compositions and methods of using galectin-8 as an inhibitor of tumor cell growth |
US7199107B2 (en) | 2002-05-23 | 2007-04-03 | Isis Pharmaceuticals, Inc. | Antisense modulation of kinesin-like 1 expression |
US20040092470A1 (en) * | 2002-06-18 | 2004-05-13 | Leonard Sherry A. | Dry powder oligonucleotide formualtion, preparation and its uses |
AU2003257181A1 (en) | 2002-08-05 | 2004-02-23 | University Of Rochester | Protein transducing domain/deaminase chimeric proteins, related compounds, and uses thereof |
WO2004016733A2 (en) | 2002-08-16 | 2004-02-26 | Agensys, Inc. | Nucleic acid and corresponding protein entitled 251p5g2 useful in treatment and detection of cancer |
CN1694959B (en) | 2002-09-13 | 2013-09-18 | 雷普利瑟公司 | Non-sequence complementary antiviral oligonucleotides |
JP2006500030A (en) * | 2002-09-20 | 2006-01-05 | イェール ユニバーシティ | Riboswitch, method of using the same, and composition for use with riboswitch |
US7229976B2 (en) | 2002-09-26 | 2007-06-12 | Isis Pharmaceuticals, Inc. | Modulation of forkhead box O1A expression |
WO2004110345A2 (en) * | 2002-10-29 | 2004-12-23 | Pharmacia Corporation | Differentially expressed genes involved in cancer, the polypeptides encoded thereby, and methods of using the same |
WO2004044132A2 (en) * | 2002-11-05 | 2004-05-27 | Isis Pharmaceuticals, Inc. | Modified oligonucleotides for use in rna interference |
US9150606B2 (en) * | 2002-11-05 | 2015-10-06 | Isis Pharmaceuticals, Inc. | Compositions comprising alternating 2'-modified nucleosides for use in gene modulation |
US9150605B2 (en) * | 2002-11-05 | 2015-10-06 | Isis Pharmaceuticals, Inc. | Compositions comprising alternating 2′-modified nucleosides for use in gene modulation |
CA2504720C (en) | 2002-11-05 | 2013-12-24 | Isis Pharmaceuticals, Inc. | Chimeric oligomeric compounds and their use in gene modulation |
EP1569695B1 (en) | 2002-11-13 | 2013-05-15 | Genzyme Corporation | Antisense modulation of apolipoprotein b expression |
DK2336318T3 (en) | 2002-11-13 | 2013-07-15 | Genzyme Corp | ANTISENSE MODULATION OF APOLIPOPROTEIN B EXPRESSION |
JP4555089B2 (en) * | 2002-11-15 | 2010-09-29 | モーフオテク・インコーポレーテツド | Method for producing high production amount of antibody from hybridoma created by in vitro immunization |
US8007804B2 (en) | 2002-11-15 | 2011-08-30 | Musc Foundation For Research Development | Complement receptor 2 targeted complement modulators |
AU2003294462C1 (en) | 2002-11-21 | 2011-06-30 | University Of Utah Research Foundation | Purinergic modulation of smell |
US7144999B2 (en) | 2002-11-23 | 2006-12-05 | Isis Pharmaceuticals, Inc. | Modulation of hypoxia-inducible factor 1 alpha expression |
US20040121338A1 (en) * | 2002-12-19 | 2004-06-24 | Alsmadi Osama A. | Real-time detection of rolling circle amplification products |
US9487823B2 (en) * | 2002-12-20 | 2016-11-08 | Qiagen Gmbh | Nucleic acid amplification |
WO2004058987A2 (en) | 2002-12-20 | 2004-07-15 | Qiagen Gmbh | Nucleic acid amplification |
JP2006516099A (en) * | 2002-12-23 | 2006-06-22 | ダイナバックス テクノロジーズ コーポレイション | Branched immunomodulatory compounds and methods of using the compounds |
US6977153B2 (en) | 2002-12-31 | 2005-12-20 | Qiagen Gmbh | Rolling circle amplification of RNA |
CA2860151A1 (en) | 2003-02-10 | 2004-08-26 | Agensys, Inc. | Nucleic acid and corresponding protein named 158p1d7 useful in the treatment and detection of bladder and other cancers |
DK1597366T3 (en) | 2003-02-11 | 2013-02-25 | Antisense Therapeutics Ltd | Modulation of expression of insulin-like growth factor receptor I |
US7002006B2 (en) * | 2003-02-12 | 2006-02-21 | Isis Pharmaceuticals, Inc. | Protection of nucleosides |
EP1592708A2 (en) | 2003-02-14 | 2005-11-09 | Sagres Discovery, Inc. | Therapeutic gpcr targets in cancer |
US20040170982A1 (en) | 2003-02-14 | 2004-09-02 | Morris David W. | Novel therapeutic targets in cancer |
US7767387B2 (en) * | 2003-06-13 | 2010-08-03 | Sagres Discovery, Inc. | Therapeutic targets in cancer |
US20070218071A1 (en) * | 2003-09-15 | 2007-09-20 | Morris David W | Novel therapeutic targets in cancer |
US6943768B2 (en) | 2003-02-21 | 2005-09-13 | Xtellus Inc. | Thermal control system for liquid crystal cell |
US7803781B2 (en) | 2003-02-28 | 2010-09-28 | Isis Pharmaceuticals, Inc. | Modulation of growth hormone receptor expression and insulin-like growth factor expression |
US20070141570A1 (en) * | 2003-03-07 | 2007-06-21 | Sequenom, Inc. | Association of polymorphic kinase anchor proteins with cardiac phenotypes and related methods |
US20040185559A1 (en) | 2003-03-21 | 2004-09-23 | Isis Pharmaceuticals Inc. | Modulation of diacylglycerol acyltransferase 1 expression |
US8043834B2 (en) | 2003-03-31 | 2011-10-25 | Qiagen Gmbh | Universal reagents for rolling circle amplification and methods of use |
US7598227B2 (en) | 2003-04-16 | 2009-10-06 | Isis Pharmaceuticals Inc. | Modulation of apolipoprotein C-III expression |
US7399853B2 (en) | 2003-04-28 | 2008-07-15 | Isis Pharmaceuticals | Modulation of glucagon receptor expression |
PT1629088E (en) | 2003-05-30 | 2012-04-10 | Agensys Inc | Prostate stem cell antigen (psca) variants and subsequences thereof |
WO2004108081A2 (en) * | 2003-06-02 | 2004-12-16 | Isis Pharmaceuticals, Inc. | Oligonucleotide synthesis with alternative solvents |
AU2004253455B2 (en) | 2003-06-03 | 2011-03-03 | Eli Lilly And Company | Modulation of survivin expression |
EP3604537B1 (en) | 2003-06-13 | 2021-12-08 | Alnylam Europe AG | Double-stranded ribonucleic acid with increased effectiveness in an organism |
US20040259100A1 (en) | 2003-06-20 | 2004-12-23 | Illumina, Inc. | Methods and compositions for whole genome amplification and genotyping |
EP1636342A4 (en) * | 2003-06-20 | 2008-10-08 | Isis Pharmaceuticals Inc | Oligomeric compounds for use in gene modulation |
US7790691B2 (en) * | 2003-06-20 | 2010-09-07 | Isis Pharmaceuticals, Inc. | Double stranded compositions comprising a 3′-endo modified strand for use in gene modulation |
US7683036B2 (en) | 2003-07-31 | 2010-03-23 | Regulus Therapeutics Inc. | Oligomeric compounds and compositions for use in modulation of small non-coding RNAs |
US7825235B2 (en) | 2003-08-18 | 2010-11-02 | Isis Pharmaceuticals, Inc. | Modulation of diacylglycerol acyltransferase 2 expression |
US7354706B2 (en) * | 2003-09-09 | 2008-04-08 | The Regents Of The University Of Colorado, A Body Corporate | Use of photopolymerization for amplification and detection of a molecular recognition event |
US20050053981A1 (en) * | 2003-09-09 | 2005-03-10 | Swayze Eric E. | Gapped oligomeric compounds having linked bicyclic sugar moieties at the termini |
US20090163375A1 (en) | 2003-09-09 | 2009-06-25 | Bowman Christopher N | Use of Photopolymerization for Amplification and Detection of a Molecular Recognition Event |
US20070123480A1 (en) * | 2003-09-11 | 2007-05-31 | Replicor Inc. | Oligonucleotides targeting prion diseases |
AU2004274021B2 (en) * | 2003-09-18 | 2009-08-13 | Isis Pharmaceuticals, Inc. | 4'-thionucleosides and oligomeric compounds |
AR045937A1 (en) | 2003-09-18 | 2005-11-16 | Lilly Co Eli | MODULATION OF THE EXPRESSION OF THE EIF4E EUCARY INITIATION FACTOR |
US20070281896A1 (en) * | 2003-09-30 | 2007-12-06 | Morris David W | Novel compositions and methods in cancer |
US20050191653A1 (en) | 2003-11-03 | 2005-09-01 | Freier Susan M. | Modulation of SGLT2 expression |
PT2161283E (en) | 2003-11-17 | 2014-08-29 | Genentech Inc | Compositions comprising antibodies against cd79b conjugated to a growth inhibitory agent or cytotoxic agent and methods for the treatment of tumor of hematopoietic origin |
US20050136414A1 (en) * | 2003-12-23 | 2005-06-23 | Kevin Gunderson | Methods and compositions for making locus-specific arrays |
EP1711606A2 (en) | 2004-01-20 | 2006-10-18 | Isis Pharmaceuticals, Inc. | Modulation of glucocorticoid receptor expression |
US8778900B2 (en) * | 2004-01-22 | 2014-07-15 | Isis Pharmaceuticals, Inc. | Modulation of eIF4E-BP1 expression |
US7468431B2 (en) * | 2004-01-22 | 2008-12-23 | Isis Pharmaceuticals, Inc. | Modulation of eIF4E-BP2 expression |
US8569474B2 (en) * | 2004-03-09 | 2013-10-29 | Isis Pharmaceuticals, Inc. | Double stranded constructs comprising one or more short strands hybridized to a longer strand |
EP2700720A3 (en) | 2004-03-15 | 2015-01-28 | Isis Pharmaceuticals, Inc. | Compositions and methods for optimizing cleavage of RNA by RNASE H |
US20050244869A1 (en) * | 2004-04-05 | 2005-11-03 | Brown-Driver Vickie L | Modulation of transthyretin expression |
WO2005097817A2 (en) | 2004-04-05 | 2005-10-20 | Alnylam Pharmaceuticals, Inc. | Process and reagents for oligonucleotide synthesis and purification |
US20050260755A1 (en) * | 2004-04-06 | 2005-11-24 | Isis Pharmaceuticals, Inc. | Sequential delivery of oligomeric compounds |
JP4584987B2 (en) | 2004-04-30 | 2010-11-24 | アルニラム ファーマスーティカルズ インコーポレイテッド | Oligonucleotides containing C5-modified pyrimidines |
EP2322662A1 (en) | 2004-05-14 | 2011-05-18 | Rosetta Genomics Ltd | MicroRNAs and uses thereof |
AU2005243410B2 (en) | 2004-05-14 | 2010-04-22 | Rosetta Genomics Ltd. | Micronas and uses thereof |
AU2005326817B2 (en) | 2004-05-21 | 2012-04-19 | Benaroya Research Institute | Variable lymphocyte receptors, related polypeptides and nucleic acids, and uses thereof |
JP2007538236A (en) | 2004-05-21 | 2007-12-27 | アトノミックス アクティーゼルスカブ | Surface acoustic wave sensor containing hydrogel |
WO2005118864A2 (en) | 2004-05-28 | 2005-12-15 | Agensys, Inc. | Antibodies and related molecules that bind to psca proteins |
AU2005252663B2 (en) * | 2004-06-03 | 2011-07-07 | Isis Pharmaceuticals, Inc. | Double strand compositions comprising differentially modified strands for use in gene modulation |
US8394947B2 (en) * | 2004-06-03 | 2013-03-12 | Isis Pharmaceuticals, Inc. | Positionally modified siRNA constructs |
US20060024677A1 (en) | 2004-07-20 | 2006-02-02 | Morris David W | Novel therapeutic targets in cancer |
US7427675B2 (en) * | 2004-08-23 | 2008-09-23 | Isis Pharmaceuticals, Inc. | Compounds and methods for the characterization of oligonucleotides |
US7884086B2 (en) * | 2004-09-08 | 2011-02-08 | Isis Pharmaceuticals, Inc. | Conjugates for use in hepatocyte free uptake assays |
DK1809303T3 (en) | 2004-09-23 | 2019-06-11 | Arc Medical Devices Inc | PHARMACEUTICAL COMPOSITION AND METHODS IN RELATION TO FIBROSE ADHESION OR INFLAMMATORIC DISEASE USING LOW SULPHATE FUCAN |
US8765700B2 (en) | 2004-10-20 | 2014-07-01 | Antisense Therapeutics Ltd. | Topical administrations of antisense compounds to VLA-4 for the treatment of respiratory conditions |
US20100256062A1 (en) | 2004-12-06 | 2010-10-07 | Howard Tommy E | Allelic Variants of Human Factor VIII |
AU2006223498A1 (en) | 2005-03-10 | 2006-09-21 | Genentech, Inc. | Methods and compositions for modulating vascular integrity |
US7476733B2 (en) * | 2005-03-25 | 2009-01-13 | The United States Of America As Represented By The Department Of Health And Human Services | Development of a real-time PCR assay for detection of pneumococcal DNA and diagnosis of pneumococccal disease |
AU2006230563B8 (en) | 2005-03-31 | 2010-06-17 | Agensys, Inc. | Antibodies and related molecules that bind to 161P2F10B proteins |
EP1863908B1 (en) * | 2005-04-01 | 2010-11-17 | Qiagen GmbH | Reverse transcription and amplification of rna with simultaneous degradation of dna |
EP2062591A1 (en) | 2005-04-07 | 2009-05-27 | Novartis Vaccines and Diagnostics, Inc. | CACNA1E in cancer diagnosis detection and treatment |
EP2083088A3 (en) | 2005-04-07 | 2009-10-14 | Novartis Vaccines and Diagnostics, Inc. | Cancer-related genes |
US9505867B2 (en) | 2005-05-31 | 2016-11-29 | Ecole Polytechmique Fédérale De Lausanne | Triblock copolymers for cytoplasmic delivery of gene-based drugs |
WO2006133022A2 (en) | 2005-06-03 | 2006-12-14 | The Johns Hopkins University | Compositions and methods for decreasing microrna expression for the treatment of neoplasia |
US8252756B2 (en) | 2005-06-14 | 2012-08-28 | Northwestern University | Nucleic acid functionalized nanoparticles for therapeutic applications |
US7776532B2 (en) | 2005-08-11 | 2010-08-17 | Synthetic Genomics, Inc. | Method for in vitro recombination |
CA2617693A1 (en) | 2005-08-17 | 2007-02-22 | Medexis S.A. | Composition and method for determination of ck19 expression |
JP5523705B2 (en) | 2005-08-29 | 2014-06-18 | レグルス・セラピューティクス・インコーポレイテッド | Method of using to modulate MIR-122A |
EP2338992A3 (en) | 2005-08-29 | 2011-10-12 | Regulus Therapeutics, Inc | Antisense compounds having enhanced anti-microRNA activity |
EP1762627A1 (en) | 2005-09-09 | 2007-03-14 | Qiagen GmbH | Method for the activation of a nucleic acid for performing a polymerase reaction |
US8080534B2 (en) | 2005-10-14 | 2011-12-20 | Phigenix, Inc | Targeting PAX2 for the treatment of breast cancer |
EP2402435A3 (en) | 2005-10-14 | 2012-03-28 | MUSC Foundation For Research Development | Targeting PAX2 for the induction of DEFB1-mediated tumor immunity and cancer therapy |
EP2325315B1 (en) | 2005-10-28 | 2014-05-07 | Alnylam Pharmaceuticals, Inc. | Compositions and methods for inhibiting expression of huntingtin gene |
CA2626584A1 (en) * | 2005-11-04 | 2007-05-18 | Alnylam Pharmaceuticals, Inc. | Compositions and methods for inhibiting expression of nav1.8 gene |
CA2626690A1 (en) | 2005-11-09 | 2007-05-18 | Alnylam Pharmaceuticals, Inc. | Compositions and methods for inhibiting expression of factor v leiden mutant gene |
US7807652B2 (en) | 2005-11-21 | 2010-10-05 | Isis Pharmaceuticals, Inc. | Modulation of eIF4E-BP2 expression |
US8313901B2 (en) * | 2005-12-21 | 2012-11-20 | Yale University | Methods and compositions related to the modulation of riboswitches |
EP1976567B1 (en) | 2005-12-28 | 2020-05-13 | The Scripps Research Institute | Natural antisense and non-coding rna transcripts as drug targets |
JP5425474B2 (en) | 2006-01-26 | 2014-02-26 | アイシス ファーマシューティカルズ, インコーポレーテッド | Composition and use thereof for huntingtin |
CA2640058C (en) | 2006-01-27 | 2018-04-24 | Isis Pharmaceuticals, Inc. | Oligomeric compounds and compositions for the use in modulation of micrornas |
US7569686B1 (en) | 2006-01-27 | 2009-08-04 | Isis Pharmaceuticals, Inc. | Compounds and methods for synthesis of bicyclic nucleic acid analogs |
DK2314594T3 (en) | 2006-01-27 | 2014-10-27 | Isis Pharmaceuticals Inc | 6-modified bicyclic nucleic acid analogues |
JP4984990B2 (en) | 2006-03-28 | 2012-07-25 | 富士通株式会社 | Method for producing functional molecules |
ES2544861T3 (en) | 2006-03-31 | 2015-09-04 | Alnylam Pharmaceuticals Inc. | Compositions and methods to inhibit the expression of the Eg5 gene |
MX2008014005A (en) * | 2006-05-03 | 2009-01-27 | Baltic Technology Dev Ltd | Antisense agents combining strongly bound base - modified oligonucleotide and artificial nuclease. |
EP2023939B1 (en) | 2006-05-05 | 2012-06-27 | Isis Pharmaceuticals, Inc. | Compounds and methods for modulating expression of pcsk9 |
EP2505648A1 (en) | 2006-05-05 | 2012-10-03 | Isis Pharmaceuticals, Inc. | Compounds and methods for modulating expression of PTP1B |
DE102006020885A1 (en) * | 2006-05-05 | 2007-11-08 | Qiagen Gmbh | Inserting a tag sequence into a nucleic acid comprises using an anchor oligonucleotide comprising a hybridizing anchor sequence and a nonhybridizing tag-template sequence |
US7666854B2 (en) * | 2006-05-11 | 2010-02-23 | Isis Pharmaceuticals, Inc. | Bis-modified bicyclic nucleic acid analogs |
WO2007134181A2 (en) * | 2006-05-11 | 2007-11-22 | Isis Pharmaceuticals, Inc. | 5'-modified bicyclic nucleic acid analogs |
WO2007137156A2 (en) | 2006-05-19 | 2007-11-29 | Alnylam Pharmaceuticals, Inc. | Rnai modulation of aha and therapeutic uses thereof |
US7888498B2 (en) | 2006-05-22 | 2011-02-15 | Alnylam Pharmaceuticals, Inc. | Compositions and methods for inhibiting expression of IKK-B gene |
EP2023938A4 (en) * | 2006-05-23 | 2010-11-10 | Isis Pharmaceuticals Inc | Modulation of chrebp expression |
CA2672246A1 (en) | 2006-06-30 | 2008-03-13 | Rosetta Genomics Ltd | Detecting nucleic acids |
WO2008011473A2 (en) | 2006-07-19 | 2008-01-24 | Isis Pharmaceuticals, Inc. | Compositions and their uses directed to hbxip |
EP2057284A4 (en) * | 2006-08-04 | 2011-06-29 | Isis Pharmaceuticals Inc | Compositions and methods for the modulation of jnk proteins |
CA2660572C (en) | 2006-08-11 | 2014-07-22 | University Of Medicine And Dentistry Of New Jersey | Dual-sensitizer-containing luminescent compounds, conjugates, and uses thereof |
US20100137440A1 (en) * | 2006-09-11 | 2010-06-03 | Yale University | Lysine riboswitches, structure-based compound design with lysine riboswitches, and methods and compositions for use of and with lysine riboswitches |
WO2008063758A2 (en) | 2006-10-05 | 2008-05-29 | Massachussetts Institute Of Technology | Multifunctional encoded particles for high-throughput analysis |
ATE540118T1 (en) | 2006-10-18 | 2012-01-15 | Isis Pharmaceuticals Inc | ANTISENSE COMPOUNDS |
WO2008136852A2 (en) | 2006-11-01 | 2008-11-13 | University Of Rochester | Methods and compositions related to the structure and function of apobec3g |
EP2102340A2 (en) * | 2006-11-27 | 2009-09-23 | Isis Pharmaceuticals, Inc. | Methods for treating hypercholesterolemia |
US8093222B2 (en) | 2006-11-27 | 2012-01-10 | Isis Pharmaceuticals, Inc. | Methods for treating hypercholesterolemia |
US8481506B2 (en) * | 2006-12-05 | 2013-07-09 | Rosetta Genomics, Ltd. | Nucleic acids involved in viral infection |
CA2672297A1 (en) | 2006-12-11 | 2008-06-19 | University Of Utah Research Foundation | Compositions and methods for treating pathologic angiogenesis and vascular permeability |
CN103966345A (en) | 2007-02-09 | 2014-08-06 | 西北大学 | Particles for detecting intracellular targets |
US20080241140A1 (en) * | 2007-02-12 | 2008-10-02 | Medical College Of Georgia | Gene amplification of coactivator coaa and uses thereof |
CA2679586A1 (en) * | 2007-02-27 | 2008-10-23 | Northwestern University | Molecule attachment to nanoparticles |
CN101801185A (en) | 2007-03-22 | 2010-08-11 | 耶鲁大学 | Methods and compositions related to riboswitches that control alternative splicing |
EP2905336A1 (en) | 2007-03-29 | 2015-08-12 | Alnylam Pharmaceuticals Inc. | Compositions and methods for inhibiting expression of a gene from the ebola |
CA2690281A1 (en) | 2007-05-11 | 2008-11-20 | The Johns Hopkins University | Biomarkers for melanoma |
EP2426219A1 (en) | 2007-05-29 | 2012-03-07 | Yale University | Riboswitches and methods and compositions for use of and with riboswitches |
EP2164994A4 (en) * | 2007-05-29 | 2010-07-21 | Univ Yale Inc | Methods and compositions related to riboswitches that control alternative splicing and rna processing |
CA2689923A1 (en) | 2007-05-30 | 2008-12-11 | Northwestern University | Nucleic acid functionalized nanoparticles for therapeutic applications |
US8278425B2 (en) | 2007-05-30 | 2012-10-02 | Isis Pharmaceuticals, Inc. | N-substituted-aminomethylene bridged bicyclic nucleic acid analogs |
US7807372B2 (en) * | 2007-06-04 | 2010-10-05 | Northwestern University | Screening sequence selectivity of oligonucleotide-binding molecules using nanoparticle based colorimetric assay |
US8278426B2 (en) | 2007-06-08 | 2012-10-02 | Isis Pharmaceuticals, Inc. | Carbocyclic bicyclic nucleic acid analogs |
US8278283B2 (en) * | 2007-07-05 | 2012-10-02 | Isis Pharmaceuticals, Inc. | 6-disubstituted or unsaturated bicyclic nucleic acid analogs |
US8088904B2 (en) | 2007-08-15 | 2012-01-03 | Isis Pharmaceuticals, Inc. | Tetrahydropyran nucleic acid analogs |
US8557767B2 (en) | 2007-08-28 | 2013-10-15 | Uab Research Foundation | Synthetic apolipoprotein E mimicking polypeptides and methods of use |
AU2008296478B9 (en) * | 2007-08-28 | 2015-03-19 | The Uab Research Foundation | Synthetic apolipoprotein E mimicking polypeptides and methods of use |
WO2009039466A1 (en) | 2007-09-20 | 2009-03-26 | Vanderbilt University | Free solution measurement of molecular interactions by backscattering interferometry |
WO2009039442A1 (en) * | 2007-09-21 | 2009-03-26 | California Institute Of Technology | Nfia in glial fate determination, glioma therapy and astrocytoma treatment |
AU2008324068A1 (en) * | 2007-11-05 | 2009-05-14 | Baltic Technology Development, Ltd. | Use of oligonucleotides with modified bases in hybridization of nucleic acids |
US20090137405A1 (en) * | 2007-11-16 | 2009-05-28 | Christopher Bowman | Detection of nucleic acid biomarkers using polymerization-based amplification |
US8916531B2 (en) * | 2007-11-20 | 2014-12-23 | Isis Pharmaceuticals, Inc. | Modulation of CD40 expression |
EP2617828B1 (en) | 2007-12-10 | 2014-09-24 | Alnylam Pharmaceuticals Inc. | Compositions and methods for inhibiting expression of factor VII gene |
US20110117125A1 (en) | 2008-01-02 | 2011-05-19 | Tekmira Pharmaceuticals Corporation | Compositions and methods for the delivery of nucleic acids |
WO2009100320A2 (en) * | 2008-02-07 | 2009-08-13 | Isis Pharmaceuticals, Inc. | Bicyclic cyclohexitol nucleic acid analogs |
AU2009221775B2 (en) | 2008-03-05 | 2015-05-07 | Alnylam Pharmaceuticals, Inc. | Compositions and methods for inhibiting expression of Eg5 and VEGF genes |
EP2282744B1 (en) | 2008-03-21 | 2018-01-17 | Ionis Pharmaceuticals, Inc. | Oligomeric compounds comprising tricyclic nucleosides and methods for their use |
US8846639B2 (en) * | 2008-04-04 | 2014-09-30 | Isis Pharmaceutical, Inc. | Oligomeric compounds comprising bicyclic nucleosides and having reduced toxicity |
WO2009124238A1 (en) * | 2008-04-04 | 2009-10-08 | Isis Pharmaceuticals, Inc. | Oligomeric compounds comprising neutrally linked terminal bicyclic nucleosides |
ES2554168T3 (en) | 2008-04-18 | 2015-12-16 | Baxter International Inc. | Microsphere-based composition to prevent and / or reverse newly occurring autoimmune diabetes |
US8222221B2 (en) | 2008-06-04 | 2012-07-17 | The Board Of Regents Of The University Of Texas System | Modulation of gene expression through endogenous small RNA targeting of gene promoters |
US20110119781A1 (en) * | 2008-07-15 | 2011-05-19 | Birgit Bramlage | Compositions and Methods for Inhibiting Expression of TGF-BETA Receptor Genes |
US8901095B2 (en) | 2008-07-29 | 2014-12-02 | The Board Of Regents Of The University Of Texas System | Selective inhibition of polyglutamine protein expression |
EP2323667A4 (en) * | 2008-08-07 | 2012-07-25 | Isis Pharmaceuticals Inc | Modulation of transthyretin expression for the treatment of cns related disorders |
ES2657696T3 (en) | 2008-08-25 | 2018-03-06 | Excaliard Pharmaceuticals, Inc. | Method to reduce healing during wound healing using antisense compounds targeting CTGF |
AU2009275387B2 (en) | 2008-08-25 | 2010-07-08 | Excaliard Pharmaceuticals, Inc. | Antisense oligonucleotides directed against connective tissue growth factor and uses thereof |
EP3208337A1 (en) | 2008-09-02 | 2017-08-23 | Alnylam Pharmaceuticals, Inc. | Compositions for combined inhibition of mutant egfr and il-6 expression |
EP2356129B1 (en) * | 2008-09-24 | 2013-04-03 | Isis Pharmaceuticals, Inc. | Substituted alpha-l-bicyclic nucleosides |
DK2361256T3 (en) | 2008-09-24 | 2013-07-01 | Isis Pharmaceuticals Inc | Cyclohexenyl-nucleic acid analogues |
EP3584320A1 (en) | 2008-09-25 | 2019-12-25 | Alnylam Pharmaceuticals, Inc. | Lipid formulated compositions and methods for inhibiting expression of serum amyloid a gene |
CN104119242B (en) | 2008-10-09 | 2017-07-07 | 泰米拉制药公司 | The amino lipids of improvement and the method for delivering nucleic acid |
US8334372B2 (en) | 2008-10-15 | 2012-12-18 | Isis Pharmaceuticals, Inc. | Modulation of factor 11 expression |
CN106834291B (en) | 2008-10-20 | 2020-09-29 | 阿尔尼拉姆医药品有限公司 | Compositions and methods for inhibiting expression of transthyretin |
EP2358397B1 (en) | 2008-10-24 | 2020-01-01 | Ionis Pharmaceuticals, Inc. | 5' and 2' bis-substituted nucleosides and oligomeric compounds prepared therefrom |
EP2447274B1 (en) | 2008-10-24 | 2017-10-04 | Ionis Pharmaceuticals, Inc. | Oligomeric compounds and methods |
CN102216457A (en) * | 2008-11-17 | 2011-10-12 | 霍夫曼-拉罗奇有限公司 | Compositions and methods for inhibiting expression of factor VII genes |
KR101692880B1 (en) | 2008-11-24 | 2017-01-04 | 노오쓰웨스턴 유니버시티 | Polyvalent rna-nanoparticle compositions |
EP2370582B1 (en) | 2008-12-04 | 2017-05-10 | CuRNA, Inc. | Treatment of tumor suppressor gene related diseases by inhibition of natural antisense transcript to the gene |
EP2370580B1 (en) | 2008-12-04 | 2019-09-11 | CuRNA, Inc. | Treatment of sirtuin 1 (sirt1) related diseases by inhibition of natural antisense transcript to sirtuin 1 |
ES2629630T3 (en) | 2008-12-04 | 2017-08-11 | Curna, Inc. | Treatment of diseases related to erythropoietin (EPO) by inhibiting the natural antisense transcript to EPO |
AU2009324534B2 (en) | 2008-12-10 | 2015-07-30 | Alnylam Pharmaceuticals, Inc. | GNAQ targeted dsRNA compositions and methods for inhibiting expression |
US20100233270A1 (en) | 2009-01-08 | 2010-09-16 | Northwestern University | Delivery of Oligonucleotide-Functionalized Nanoparticles |
US20100184844A1 (en) * | 2009-01-08 | 2010-07-22 | Northwestern University | Inhibition of Bacterial Protein Production by Polyvalent Oligonucleotide Modified Nanoparticle Conjugates |
KR101546673B1 (en) * | 2009-01-15 | 2015-08-25 | 삼성전자주식회사 | Toner for electrophotographic and process for preparing the same |
US20120101148A1 (en) | 2009-01-29 | 2012-04-26 | Alnylam Pharmaceuticals, Inc. | lipid formulation |
KR20110100316A (en) * | 2009-02-03 | 2011-09-09 | 에프. 호프만-라 로슈 아게 | Compositions and methods for inhibiting expression of ptp1b genes |
US8536320B2 (en) | 2009-02-06 | 2013-09-17 | Isis Pharmaceuticals, Inc. | Tetrahydropyran nucleic acid analogs |
WO2010091308A2 (en) | 2009-02-06 | 2010-08-12 | Isis Pharmaceuticals, Inc. | Oligomeric compounds and methods |
ES2560107T3 (en) | 2009-02-12 | 2016-02-17 | Curna, Inc. | Treatment of diseases related to brain-derived neurotrophic factor (BDNF) by inhibition of natural antisense transcript for BDNF |
WO2010093906A2 (en) | 2009-02-12 | 2010-08-19 | Curna, Inc. | Treatment of glial cell derived neurotrophic factor (gdnf) related diseases by inhibition of natural antisense transcript to gdnf |
EP2399129B1 (en) | 2009-02-20 | 2015-11-25 | Michael P. Lisanti | A method of diagnosis or prognosis of a neoplasm comprising determining the level of expression of a protein in stromal cells adjacent to the neoplasm |
US20120041051A1 (en) | 2009-02-26 | 2012-02-16 | Kevin Fitzgerald | Compositions And Methods For Inhibiting Expression Of MIG-12 Gene |
WO2010102058A2 (en) | 2009-03-04 | 2010-09-10 | Curna, Inc. | Treatment of sirtuin 1 (sirt1) related diseases by inhibition of natural antisense transcript to sirt 1 |
WO2010105209A1 (en) | 2009-03-12 | 2010-09-16 | Alnylam Pharmaceuticals, Inc. | LIPID FORMULATED COMPOSITIONS AND METHODS FOR INHIBITING EXPRESSION OF Eg5 AND VEGF GENES |
CA2755409C (en) | 2009-03-16 | 2019-04-30 | Joseph Collard | Treatment of nuclear factor (erythroid-derived 2)-like 2 (nrf2) related diseases by inhibition of natural antisense transcript to nrf2 |
JP5904935B2 (en) | 2009-03-17 | 2016-04-20 | クルナ・インコーポレーテッド | Treatment of DLK1-related diseases by suppression of natural antisense transcripts against Delta-like 1 homolog (DLK1) |
AU2010237001B2 (en) | 2009-04-15 | 2016-07-07 | Northwestern University | Delivery of oligonucleotide-functionalized nanoparticles |
EP3248618A1 (en) | 2009-04-22 | 2017-11-29 | Massachusetts Institute Of Technology | Innate immune suppression enables repeated delivery of long rna molecules |
WO2010124231A2 (en) | 2009-04-24 | 2010-10-28 | The Board Of Regents Of The University Of Texas System | Modulation of gene expression using oligomers that target gene regions downstream of 3' untranslated regions |
CA2760589C (en) | 2009-05-01 | 2019-08-20 | Joseph Collard | Treatment of hemoglobin (hbf/hbg) related diseases by inhibition of natural antisense transcript to hbf/hbg |
KR20180094137A (en) | 2009-05-05 | 2018-08-22 | 알닐람 파마슈티칼스 인코포레이티드 | Lipid compositions |
CA3045126A1 (en) | 2009-05-05 | 2010-11-11 | Arbutus Biopharma Corporation | Methods of delivering oligonucleotides to immune cells |
KR101722541B1 (en) | 2009-05-06 | 2017-04-04 | 큐알엔에이, 인크. | Treatment of tristetraproline(ttp) related diseases by inhibition of natural antisense transcript to ttp |
CN102459596B (en) | 2009-05-06 | 2016-09-07 | 库尔纳公司 | By suppression therapy lipid transfer and the metabolic gene relevant disease of the natural antisense transcript for lipid transfer and metabolic gene |
WO2010132665A1 (en) | 2009-05-15 | 2010-11-18 | Yale University | Gemm riboswitches, structure-based compound design with gemm riboswitches, and methods and compositions for use of and with gemm riboswitches |
CA2759838A1 (en) * | 2009-05-15 | 2010-11-18 | F. Hoffmann-La Roche Ag | Compositions and methods for inhibiting expression of glucocorticoid receptor (gcr) genes |
ES2664590T3 (en) | 2009-05-18 | 2018-04-20 | Curna, Inc. | Treatment of diseases related to reprogramming factors by inhibition of the natural antisense transcript to a reprogramming factor |
US8895527B2 (en) | 2009-05-22 | 2014-11-25 | Curna, Inc. | Treatment of transcription factor E3 (TFE3) and insulin receptor substrate 2(IRS2) related diseases by inhibition of natural antisense transcript to TFE3 |
WO2010138806A2 (en) | 2009-05-28 | 2010-12-02 | Curna, Inc. | Treatment of antiviral gene related diseases by inhibition of natural antisense transcript to an antiviral gene |
US8158601B2 (en) | 2009-06-10 | 2012-04-17 | Alnylam Pharmaceuticals, Inc. | Lipid formulation |
JP5944311B2 (en) | 2009-06-16 | 2016-07-05 | クルナ・インコーポレーテッド | Treatment of collagen gene-related diseases by suppression of natural antisense transcripts against collagen genes |
US8951981B2 (en) | 2009-06-16 | 2015-02-10 | Curna, Inc. | Treatment of paraoxonase 1 (PON1) related diseases by inhibition of natural antisense transcript to PON1 |
SI3305302T1 (en) | 2009-06-17 | 2018-12-31 | Biogen Ma Inc. | Compositions and methods for modulation of smn2 splicing in a subject |
JP6073133B2 (en) | 2009-06-24 | 2017-02-01 | クルナ・インコーポレーテッド | Treatment of TNFR2-related diseases by suppression of natural antisense transcripts against tumor necrosis factor receptor 2 (TNFR2) |
EP2446037B1 (en) | 2009-06-26 | 2016-04-20 | CuRNA, Inc. | Treatment of down syndrome gene related diseases by inhibition of natural antisense transcript to a down syndrome gene |
CN102762731B (en) | 2009-08-05 | 2018-06-22 | 库尔纳公司 | By inhibiting to treat insulin gene (INS) relevant disease for the natural antisense transcript of insulin gene (INS) |
EP2462153B1 (en) | 2009-08-06 | 2015-07-29 | Isis Pharmaceuticals, Inc. | Bicyclic cyclohexose nucleic acid analogs |
WO2011020023A2 (en) | 2009-08-14 | 2011-02-17 | Alnylam Pharmaceuticals, Inc. | Lipid formulated compositions and methods for inhibiting expression of a gene from the ebola virus |
WO2011022420A1 (en) | 2009-08-17 | 2011-02-24 | Yale University | Methylation biomarkers and methods of use |
WO2011031482A2 (en) | 2009-08-25 | 2011-03-17 | Curna, Inc. | Treatment of 'iq motif containing gtpase activating protein' (iqgap) related diseases by inhibition of natural antisense transcript to iqgap |
ES2599076T3 (en) | 2009-09-02 | 2017-01-31 | Genentech, Inc. | Smoothened mutant and methods of use thereof |
KR20120105446A (en) | 2009-10-22 | 2012-09-25 | 제넨테크, 인크. | Methods and compositions for modulating hepsin activation of macrophage-stimulating protein |
JP6147502B2 (en) | 2009-10-27 | 2017-06-14 | スウィフト バイオサイエンシーズ, インコーポレイテッド | Polynucleotide primers and probes |
KR20120136345A (en) | 2009-10-30 | 2012-12-18 | 노오쓰웨스턴 유니버시티 | Templated nanoconjugates |
US20110110860A1 (en) | 2009-11-02 | 2011-05-12 | The Board Of Regents Of The University Of Texas System | Modulation of ldl receptor gene expression with double-stranded rnas targeting the ldl receptor gene promoter |
WO2011056215A1 (en) | 2009-11-03 | 2011-05-12 | Landers James P | Versatile, visible method for detecting polymeric analytes |
WO2011054939A2 (en) | 2009-11-09 | 2011-05-12 | F. Hoffmann-La Roche Ag | Compositions and methods for inhibiting expression of kif10 genes |
JP2013511285A (en) | 2009-11-23 | 2013-04-04 | スイフト・バイオサイエンシズ・インコーポレイテツド | Device for extending single-stranded target molecules |
AU2010324686B2 (en) | 2009-11-30 | 2016-05-19 | Genentech, Inc. | Antibodies for treating and diagnosing tumors expressing SLC34A2 (TAT211 = SEQID2 ) |
ES2661813T3 (en) | 2009-12-16 | 2018-04-04 | Curna, Inc. | Treatment of diseases related to membrane transcription factor peptidase, site 1 (mbtps1) by inhibition of the natural antisense transcript to the mbtps1 gene |
US20110152349A1 (en) | 2009-12-18 | 2011-06-23 | Anke Geick | Compositions and methods for inhibiting expression of il-18 genes |
JP5934106B2 (en) | 2009-12-23 | 2016-06-15 | カッパーアールエヌエー,インコーポレイテッド | Treatment of HGF-related diseases by inhibition of natural antisense transcripts against hepatocyte growth factor (HGF) |
CA2782375C (en) | 2009-12-23 | 2023-10-31 | Opko Curna, Llc | Treatment of uncoupling protein 2 (ucp2) related diseases by inhibition of natural antisense transcript to ucp2 |
EP2519634B1 (en) | 2009-12-29 | 2016-06-01 | CuRNA, Inc. | TREATMENT OF TUMOR PROTEIN 63 (p63) RELATED DISEASES BY INHIBITION OF NATURAL ANTISENSE TRANSCRIPT TO p63 |
US8921334B2 (en) | 2009-12-29 | 2014-12-30 | Curna, Inc. | Treatment of nuclear respiratory factor 1 (NRF1) related diseases by inhibition of natural antisense transcript to NRF1 |
KR101878501B1 (en) | 2010-01-04 | 2018-08-07 | 큐알엔에이, 인크. | Treatment of interferon regulatory factor 8 (irf8) related diseases by inhibition of natural antisense transcript to irf8 |
KR101853509B1 (en) | 2010-01-06 | 2018-04-30 | 큐알엔에이, 인크. | Treatment of Pancreatic Developmental Gene Related Diseases By Inhibition of Natural Antisense Transcript to A Pancreatic Developmental Gene |
WO2011085102A1 (en) | 2010-01-11 | 2011-07-14 | Isis Pharmaceuticals, Inc. | Base modified bicyclic nucleosides and oligomeric compounds prepared therefrom |
ES2664866T3 (en) | 2010-01-11 | 2018-04-23 | Curna, Inc. | Treatment of diseases related to sex hormone binding globulin (shbg) by inhibition of the natural antisense transcript to shbg |
SG182365A1 (en) | 2010-01-12 | 2012-08-30 | Univ Yale | Structured rna motifs and compounds and methods for their use |
US9221759B2 (en) | 2010-01-13 | 2015-12-29 | Rutgers, The State University Of New Jersey | Fluorophore chelated lanthanide luminescent probes with improved quantum efficiency |
WO2011088391A2 (en) * | 2010-01-14 | 2011-07-21 | Haplomics, Inc. | Predicting and reducing alloimmunogenicity of protein therapeutics |
CN102782135A (en) | 2010-01-25 | 2012-11-14 | 库尔纳公司 | Treatment of RNase H1 related diseases by inhibition of natural antisense transcript to RNase H1 |
US9574191B2 (en) | 2010-02-03 | 2017-02-21 | The Board Of Regents Of The University Of Texas System | Selective inhibition of polyglutamine protein expression |
US20130028889A1 (en) | 2010-02-04 | 2013-01-31 | Ico Therapeutics Inc. | Dosing regimens for treating and preventing ocular disorders using c-raf antisense |
US20110196016A1 (en) * | 2010-02-05 | 2011-08-11 | Anke Geick | Compositions and Methods for Inhibiting Expression of IKK2 Genes |
US8962586B2 (en) | 2010-02-22 | 2015-02-24 | Curna, Inc. | Treatment of pyrroline-5-carboxylate reductase 1 (PYCR1) related diseases by inhibition of natural antisense transcript to PYCR1 |
WO2011105902A2 (en) | 2010-02-23 | 2011-09-01 | Academisch Ziekenhuis Bij De Universiteit Van Amsterdam | Antagonists of complement component 8-beta (c8-beta) and uses thereof |
WO2011105900A2 (en) | 2010-02-23 | 2011-09-01 | Academisch Ziekenhuis Bij De Universiteit Van Amsterdam | Antagonists of complement component 8-alpha (c8-alpha) and uses thereof |
WO2011105901A2 (en) | 2010-02-23 | 2011-09-01 | Academisch Ziekenhuis Bij De Universiteit Van Amsterdam | Antagonists of complement component 9 (c9) and uses thereof |
US8877897B2 (en) | 2010-02-23 | 2014-11-04 | Genentech, Inc. | Compositions and methods for the diagnosis and treatment of tumor |
WO2011112516A1 (en) | 2010-03-08 | 2011-09-15 | Ico Therapeutics Inc. | Treating and preventing hepatitis c virus infection using c-raf kinase antisense oligonucleotides |
US20130101512A1 (en) | 2010-03-12 | 2013-04-25 | Chad A. Mirkin | Crosslinked polynucleotide structure |
WO2011115818A1 (en) | 2010-03-17 | 2011-09-22 | Isis Pharmaceuticals, Inc. | 5'-substituted bicyclic nucleosides and oligomeric compounds prepared therefrom |
JP5860029B2 (en) | 2010-03-29 | 2016-02-16 | アルナイラム ファーマシューティカルズ, インコーポレイテッドAlnylam Pharmaceuticals, Inc. | SiRNA therapy for transthyretin (TTR) related ocular amyloidosis |
CN102858979B (en) | 2010-04-09 | 2018-01-26 | 库尔纳公司 | FGF21 relevant diseases are treated by suppressing the natural antisense transcript of FGF2 1 (FGF21) |
WO2011133695A2 (en) | 2010-04-20 | 2011-10-27 | Swift Biosciences, Inc. | Materials and methods for nucleic acid fractionation by solid phase entrapment and enzyme-mediated detachment |
CN103154014B (en) | 2010-04-28 | 2015-03-25 | Isis制药公司 | Modified nucleosides, modified nucleosides-like and oligomeric compounds prepared therefrom |
US9127033B2 (en) | 2010-04-28 | 2015-09-08 | Isis Pharmaceuticals, Inc. | 5′ modified nucleosides and oligomeric compounds prepared therefrom |
SI2563920T1 (en) | 2010-04-29 | 2017-05-31 | Ionis Pharmaceuticals, Inc. | Modulation of transthyretin expression |
WO2011139387A1 (en) | 2010-05-03 | 2011-11-10 | Opko Curna, Llc | Treatment of sirtuin (sirt) related diseases by inhibition of natural antisense transcript to a sirtuin (sirt) |
BR112012028010A2 (en) | 2010-05-03 | 2017-09-26 | Genentech Inc | isolated antibody, cell, isolated nucleic acid, method of identifying a first antibody that binds to a tat425 antigenic epitope attached to an antibody, methods of inhibiting cell growth, therapeutic treatment of determining the presence of a tat425 protein and diagnosing the presence of a tumor in a mammal |
TWI586356B (en) | 2010-05-14 | 2017-06-11 | 可娜公司 | Treatment of par4 related diseases by inhibition of natural antisense transcript to par4 |
WO2011150226A1 (en) | 2010-05-26 | 2011-12-01 | Landers James P | Method for detecting nucleic acids based on aggregate formation |
NO2576783T3 (en) | 2010-05-26 | 2018-04-28 | ||
CA2801066C (en) | 2010-06-02 | 2021-02-09 | Alnylam Pharmaceuticals, Inc. | Compositions and methods directed to treating liver fibrosis |
WO2011156434A2 (en) | 2010-06-07 | 2011-12-15 | Firefly Bioworks, Inc. | Nucleic acid detection and quantification by post-hybridization labeling and universal encoding |
US8957200B2 (en) | 2010-06-07 | 2015-02-17 | Isis Pharmaceuticals, Inc. | Bicyclic nucleosides and oligomeric compounds prepared therefrom |
EP2580228B1 (en) | 2010-06-08 | 2016-03-23 | Ionis Pharmaceuticals, Inc. | Substituted 2'-amino and 2'-thio-bicyclic nucleosides and oligomeric compounds prepared therefrom |
US9638632B2 (en) | 2010-06-11 | 2017-05-02 | Vanderbilt University | Multiplexed interferometric detection system and method |
WO2011163466A1 (en) | 2010-06-23 | 2011-12-29 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | Regulation of skin pigmentation by neuregulin-1 (nrg-1) |
US8980860B2 (en) | 2010-07-14 | 2015-03-17 | Curna, Inc. | Treatment of discs large homolog (DLG) related diseases by inhibition of natural antisense transcript to DLG |
US20130237585A1 (en) | 2010-07-19 | 2013-09-12 | University Of Rochester | Modulation of dystrophia myotonica-protein kinase (dmpk) expression |
WO2012021554A1 (en) | 2010-08-09 | 2012-02-16 | Yale University | Cyclic di-gmp-ii riboswitches, motifs, and compounds, and methods for their use |
DK2625197T3 (en) | 2010-10-05 | 2016-10-03 | Genentech Inc | Smoothened MUTANT AND METHODS OF USING THE SAME |
US8993533B2 (en) | 2010-10-06 | 2015-03-31 | Curna, Inc. | Treatment of sialidase 4 (NEU4) related diseases by inhibition of natural antisense transcript to NEU4 |
WO2012052258A1 (en) | 2010-10-18 | 2012-04-26 | Arrowhead Research Corporation | Compositions and methods for inhibiting expression of rrm2 genes |
CA2815212A1 (en) | 2010-10-22 | 2012-04-26 | Curna, Inc. | Treatment of alpha-l-iduronidase (idua) related diseases by inhibition of natural antisense transcript to idua |
DK2633052T3 (en) | 2010-10-27 | 2018-07-16 | Curna Inc | TREATMENT OF INTERFERON-RELATED DEVELOPMENT REGULATOR 1 (IFRD1) -RELATED DISEASES BY INHIBITION OF NATURAL ANTISENCE TRANSCRIPT TO IFRD1 |
WO2012064824A1 (en) | 2010-11-09 | 2012-05-18 | Alnylam Pharmaceuticals, Inc. | Lipid formulated compositions and methods for inhibiting expression of eg5 and vegf genes |
CA3077910A1 (en) | 2010-11-17 | 2012-05-24 | Ionis Pharmaceuticals, Inc. | Modulation of alpha synuclein expression |
KR102010598B1 (en) | 2010-11-23 | 2019-08-13 | 큐알엔에이, 인크. | Treatment of nanog related diseases by inhibition of natural antisense transcript to nanog |
US9150926B2 (en) | 2010-12-06 | 2015-10-06 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | Diagnosis and treatment of adrenocortical tumors using human microRNA-483 |
WO2012078967A2 (en) | 2010-12-10 | 2012-06-14 | Alnylam Pharmaceuticals, Inc. | Compositions and methods for increasing erythropoietin (epo) production |
WO2012079046A2 (en) | 2010-12-10 | 2012-06-14 | Alnylam Pharmaceuticals, Inc. | Compositions and methods for inhibiting expression of klf-1 and bcl11a genes |
WO2012106509A1 (en) | 2011-02-02 | 2012-08-09 | The Trustees Of Princeton University | Sirtuin modulators as virus production modulators |
WO2012106508A1 (en) | 2011-02-02 | 2012-08-09 | Pfizer Inc. | Method of treating keloids or hypertrophic scars using antisense compounds targeting connective tissue growth factor (ctgf) |
BR112013019853A2 (en) | 2011-02-03 | 2017-03-21 | Mirna Therapeutics Inc | mir-34 synthetic mimetics |
CN105969773A (en) | 2011-02-03 | 2016-09-28 | 米尔纳医疗股份有限公司 | Synthetic mimics of MIR-124 |
EP3467109A1 (en) | 2011-02-08 | 2019-04-10 | Ionis Pharmaceuticals, Inc. | Oligomeric compounds comprising bicyclic nucleotides and uses thereof |
US9562853B2 (en) | 2011-02-22 | 2017-02-07 | Vanderbilt University | Nonaqueous backscattering interferometric methods |
SG193923A1 (en) | 2011-03-29 | 2013-11-29 | Alnylam Pharmaceuticals Inc | Compositions and methods for inhibiting expression of tmprss6 gene |
EP3460064B8 (en) | 2011-04-03 | 2024-03-20 | The General Hospital Corporation d/b/a Massachusetts General Hospital | Efficient protein expression in vivo using modified rna (mod-rna) |
WO2012149154A1 (en) | 2011-04-26 | 2012-11-01 | Swift Biosciences, Inc. | Polynucleotide primers and probes |
WO2012151289A2 (en) | 2011-05-02 | 2012-11-08 | University Of Virginia Patent Foundation | Method and system to detect aggregate formation on a substrate |
WO2012151268A1 (en) | 2011-05-02 | 2012-11-08 | University Of Virginia Patent Foundation | Method and system for high throughput optical and label free detection of analytes |
WO2012170347A1 (en) | 2011-06-09 | 2012-12-13 | Isis Pharmaceuticals, Inc. | Bicyclic nucleosides and oligomeric compounds prepared therefrom |
JP6188686B2 (en) | 2011-06-09 | 2017-08-30 | カッパーアールエヌエー,インコーポレイテッド | Treatment of FXN-related diseases by inhibition of natural antisense transcripts to frataxin (FXN) |
WO2012170945A2 (en) | 2011-06-10 | 2012-12-13 | Isis Pharmaceuticals, Inc. | Methods for modulating kallikrein (klkb1) expression |
SG10201800715PA (en) | 2011-06-21 | 2018-02-27 | Alnylam Pharmaceuticals Inc | Angiopoietin-like 3 (angptl3) irna compostions and methods of use thereof |
EP2723351B1 (en) | 2011-06-21 | 2018-02-14 | Alnylam Pharmaceuticals, Inc. | Compositions and methods for inhibition of expression of protein c (proc) genes |
MX344807B (en) | 2011-06-21 | 2017-01-09 | Alnylam Pharmaceuticals Inc | Compositions and methods for inhibition of expression of apolipoprotein c-iii (apoc3) genes. |
WO2012178033A2 (en) | 2011-06-23 | 2012-12-27 | Alnylam Pharmaceuticals, Inc. | Serpina1 sirnas: compositions of matter and methods of treatment |
CA2840614A1 (en) | 2011-06-29 | 2013-01-03 | Isis Pharmaceuticals, Inc. | Methods for modulating kallikrein (klkb1) expression |
DK2726613T3 (en) | 2011-06-30 | 2018-12-03 | Arrowhead Pharmaceuticals Inc | COMPOSITIONS AND PROCEDURES FOR INHIBITING GENEPRESSION OF HEPATITIS B VIRUS |
EP2739735A2 (en) | 2011-08-01 | 2014-06-11 | Alnylam Pharmaceuticals, Inc. | Method for improving the success rate of hematopoietic stem cell transplants |
EP2742135B2 (en) | 2011-08-11 | 2020-06-10 | Ionis Pharmaceuticals, Inc. | Linkage modified gapped oligomeric compounds and uses thereof |
AU2012308302A1 (en) | 2011-09-14 | 2014-03-20 | Northwestern University | Nanoconjugates able to cross the blood-brain barrier |
WO2013040548A2 (en) | 2011-09-17 | 2013-03-21 | Yale University | Fluoride-responsive riboswitchs, fluoride transporters, and methods of use |
MX2014006130A (en) | 2011-11-22 | 2015-04-13 | Intermune Inc | Methods of diagnosing and treating idiopathic pulmonary fibrosis. |
CN104114572A (en) | 2011-12-16 | 2014-10-22 | 现代治疗公司 | Modified nucleoside, nucleotide, and nucleic acid compositions |
WO2013096837A1 (en) | 2011-12-22 | 2013-06-27 | Isis Pharmaceuticals, Inc. | Methods for modulating metastasis-associated-in-lung-adenocarcinoma-transcript-1(malat-1) expression |
WO2013106358A1 (en) | 2012-01-10 | 2013-07-18 | Hussain M Mahmood | Method of treating hyperlipidemia and atherosclerosis with mir-30c |
US10085987B2 (en) | 2012-01-27 | 2018-10-02 | Thomas Jefferson University | MCT protein inhibitor-related prognostic and therapeutic methods |
WO2013120003A1 (en) | 2012-02-08 | 2013-08-15 | Isis Pharmaceuticals, Inc. | Modulation of rna by repeat targeting |
CN104395480B (en) | 2012-03-13 | 2018-01-30 | 斯威夫特生物科学公司 | For the method and composition of size-controlled homopolymeric tailing to be carried out to substrate polynucleotide by nucleic acid polymerase |
CN110438125A (en) | 2012-03-15 | 2019-11-12 | 科纳公司 | By inhibiting the natural antisense transcript of brain derived neurotrophic factor (BDNF) to treat BDNF related disease |
CA2907072A1 (en) | 2012-03-16 | 2013-09-19 | Valerion Therapeutics, Llc | Antisense conjugates for decreasing expression of dmpk |
WO2013142514A1 (en) | 2012-03-19 | 2013-09-26 | Isis Pharmaceuticals, Inc. | Methods and compositions for modulating alpha-1-antitrypsin expression |
EP2833920A2 (en) | 2012-04-02 | 2015-02-11 | Moderna Therapeutics, Inc. | Modified polynucleotides for the production of biologics and proteins associated with human disease |
AU2013243949A1 (en) | 2012-04-02 | 2014-10-30 | Moderna Therapeutics, Inc. | Modified polynucleotides for the production of biologics and proteins associated with human disease |
WO2013154799A1 (en) | 2012-04-09 | 2013-10-17 | Isis Pharmaceuticals, Inc. | Tricyclic nucleosides and oligomeric compounds prepared therefrom |
EP2850092B1 (en) | 2012-04-09 | 2017-03-01 | Ionis Pharmaceuticals, Inc. | Tricyclic nucleic acid analogs |
US9133461B2 (en) | 2012-04-10 | 2015-09-15 | Alnylam Pharmaceuticals, Inc. | Compositions and methods for inhibiting expression of the ALAS1 gene |
EP2839006B1 (en) | 2012-04-20 | 2018-01-03 | Ionis Pharmaceuticals, Inc. | Oligomeric compounds comprising bicyclic nucleotides and uses thereof |
US9127274B2 (en) | 2012-04-26 | 2015-09-08 | Alnylam Pharmaceuticals, Inc. | Serpinc1 iRNA compositions and methods of use thereof |
US9273949B2 (en) | 2012-05-11 | 2016-03-01 | Vanderbilt University | Backscattering interferometric methods |
WO2013177248A2 (en) | 2012-05-22 | 2013-11-28 | Isis Pharmaceuticals, Inc. | Modulation of enhancer rna mediated gene expression |
LT3461895T (en) | 2012-06-25 | 2020-09-10 | Ionis Pharmaceuticals, Inc. | Modulation of ube3a-ats expression |
US20140038182A1 (en) | 2012-07-17 | 2014-02-06 | Dna Logix, Inc. | Cooperative primers, probes, and applications thereof |
US20150297629A1 (en) | 2012-07-27 | 2015-10-22 | Isis Pharmaceuticals, Inc. | Modulation of renin-angiotensin system (ras) related diseases by angiotensinogen |
EP2885312A4 (en) | 2012-08-15 | 2016-01-20 | Isis Pharmaceuticals Inc | Method of preparing oligomeric compounds using modified capping protocols |
EP2897633B1 (en) | 2012-09-18 | 2020-01-01 | UTI Limited Partnership | Treatment of pain by inhibition of usp5 de-ubiquitinase |
US9175291B2 (en) | 2012-10-11 | 2015-11-03 | Isis Pharmaceuticals Inc. | Modulation of androgen receptor expression |
WO2014059353A2 (en) | 2012-10-11 | 2014-04-17 | Isis Pharmaceuticals, Inc. | Oligomeric compounds comprising bicyclic nucleosides and uses thereof |
WO2014059364A1 (en) | 2012-10-11 | 2014-04-17 | Isis Pharmaceuticals, Inc. | Methods of treating kennedy's disease |
US9029335B2 (en) | 2012-10-16 | 2015-05-12 | Isis Pharmaceuticals, Inc. | Substituted 2′-thio-bicyclic nucleosides and oligomeric compounds prepared therefrom |
CA3201145A1 (en) | 2012-10-26 | 2014-05-01 | Geron Corporation | C-myc antisense oligonucleotides and methods for using the same to treat cell-proliferative disorders |
CA2890207A1 (en) | 2012-11-05 | 2014-05-08 | Foundation Medicine, Inc. | Novel ntrk1 fusion molecules and uses thereof |
WO2014074785A1 (en) | 2012-11-08 | 2014-05-15 | Ludwig Institute For Cancer Research Ltd. | Methods of predicting outcome and treating breast cancer |
PL2922554T3 (en) | 2012-11-26 | 2022-06-20 | Modernatx, Inc. | Terminally modified rna |
BR112015013311A2 (en) | 2012-12-07 | 2017-11-14 | Haplomics Inc | tolerance induction and factor 8 mutation repair |
EP2946014A2 (en) | 2013-01-17 | 2015-11-25 | Moderna Therapeutics, Inc. | Signal-sensor polynucleotides for the alteration of cellular phenotypes |
US10980804B2 (en) | 2013-01-18 | 2021-04-20 | Foundation Medicine, Inc. | Methods of treating cholangiocarcinoma |
KR102190852B1 (en) | 2013-01-31 | 2020-12-14 | 아이오니스 파마수티컬즈, 인코포레이티드 | Method of preparing oligomeric compounds using modified coupling protocols |
KR102169899B1 (en) | 2013-02-14 | 2020-10-26 | 아이오니스 파마수티컬즈, 인코포레이티드 | Modulation of apolipoprotein c-iii (apociii) expression in lipoprotein lipase deficient (lpld) populations |
WO2014130922A1 (en) | 2013-02-25 | 2014-08-28 | Trustees Of Boston University | Compositions and methods for treating fungal infections |
US10398661B2 (en) | 2013-02-28 | 2019-09-03 | The Board Of Regents Of The University Of Texas System | Methods for classifying a cancer as susceptible to TMEPAI-directed therapies and treating such cancers |
US20160024181A1 (en) | 2013-03-13 | 2016-01-28 | Moderna Therapeutics, Inc. | Long-lived polynucleotide molecules |
WO2014160129A2 (en) | 2013-03-14 | 2014-10-02 | Alnylam Pharmaceuticals, Inc. | Complement component c5 irna compositions and methods of use thereof |
US10258698B2 (en) | 2013-03-14 | 2019-04-16 | Modernatx, Inc. | Formulation and delivery of modified nucleoside, nucleotide, and nucleic acid compositions |
US8980864B2 (en) | 2013-03-15 | 2015-03-17 | Moderna Therapeutics, Inc. | Compositions and methods of altering cholesterol levels |
JP2016522679A (en) | 2013-04-04 | 2016-08-04 | プレジデント アンド フェローズ オブ ハーバード カレッジ | Therapeutic use of genome editing with the CRISPR / Cas system |
WO2014172698A1 (en) | 2013-04-19 | 2014-10-23 | Isis Pharmaceuticals, Inc. | Compositions and methods for modulation nucleic acids through nonsense mediated decay |
US10022372B2 (en) | 2013-04-19 | 2018-07-17 | Thomas Jefferson University | Caveolin-1 related methods for treating glioblastoma with temozolomide |
US9127276B2 (en) | 2013-05-01 | 2015-09-08 | Isis Pharmaceuticals, Inc. | Conjugated antisense compounds and their use |
CN105452463B (en) | 2013-05-22 | 2019-06-21 | 阿尔尼拉姆医药品有限公司 | TMPRSS6 IRNA composition and its application method |
BR112015029139B1 (en) | 2013-05-22 | 2022-07-12 | Alnylam Pharmaceuticals, Inc | DOUBLE-STRAND RNAI AGENT FOR INHIBITING SERPINA1 EXPRESSION IN A CELL, ITS USES, AS WELL AS PHARMACEUTICAL COMPOSITION AND IN VITRO METHOD OF INHIBITING SERPINA1 EXPRESSION IN A CELL |
US20160113911A1 (en) | 2013-06-06 | 2016-04-28 | The General Hospital Corporation | Methods and compositions for the treatment of cancer |
JP6869720B2 (en) | 2013-06-13 | 2021-05-12 | アンチセンス セラピューティクス リミテッド | Combination therapy |
EP3656386A1 (en) | 2013-06-21 | 2020-05-27 | Ionis Pharmaceuticals, Inc. | Compounds and methods for modulating apolipoprotein c-iii expression for improving a diabetic profile |
WO2015002971A2 (en) | 2013-07-02 | 2015-01-08 | Isis Pharmaceuticals, Inc. | Modulators of growth hormone receptor |
AU2014287009B2 (en) | 2013-07-11 | 2020-10-29 | Modernatx, Inc. | Compositions comprising synthetic polynucleotides encoding CRISPR related proteins and synthetic sgRNAs and methods of use |
EP4079745A1 (en) | 2013-08-08 | 2022-10-26 | The Scripps Research Institute | A method for the site-specific enzymatic labelling of nucleic acids in vitro by incorporation of unnatural nucleotides |
TW201536329A (en) | 2013-08-09 | 2015-10-01 | Isis Pharmaceuticals Inc | Compounds and methods for modulation of dystrophia myotonica-protein kinase (DMPK) expression |
CA2921839A1 (en) | 2013-08-28 | 2015-03-05 | Ionis Pharmaceuticals, Inc. | Modulation of prekallikrein (pkk) expression |
AU2014315287A1 (en) | 2013-09-03 | 2015-03-12 | Moderna Therapeutics, Inc. | Chimeric polynucleotides |
US20160194368A1 (en) | 2013-09-03 | 2016-07-07 | Moderna Therapeutics, Inc. | Circular polynucleotides |
PL3043827T3 (en) | 2013-09-13 | 2020-03-31 | Ionis Pharmaceuticals, Inc. | Modulators of complement factor b |
MX2016004230A (en) | 2013-10-02 | 2016-10-21 | Alnylam Pharmaceuticals Inc | Compositions and methods for inhibiting expression of the lect2 gene. |
EP3052521A1 (en) | 2013-10-03 | 2016-08-10 | Moderna Therapeutics, Inc. | Polynucleotides encoding low density lipoprotein receptor |
WO2015051366A2 (en) | 2013-10-04 | 2015-04-09 | Novartis Ag | Novel formats for organic compounds for use in rna interference |
SG11201602631XA (en) | 2013-10-04 | 2016-05-30 | Alnylam Pharmaceuticals Inc | Compositions and methods for inhibiting expression of the alas1 gene |
JP6694811B2 (en) | 2013-10-04 | 2020-05-20 | ノバルティス アーゲー | 3'end cap for RNAi agents for use in RNA interference |
JP6546161B2 (en) | 2013-10-04 | 2019-07-17 | ノバルティス アーゲー | Organic compounds for treating hepatitis B virus |
EP3055426B1 (en) | 2013-10-09 | 2019-06-19 | The United States of America as represented by The Secretary Department of Health and Human Services | Detection of hepatitis delta virus (hdv) for the diagnosis and treatment of sjögren's syndrome and lymphoma |
US11162096B2 (en) | 2013-10-14 | 2021-11-02 | Ionis Pharmaceuticals, Inc | Methods for modulating expression of C9ORF72 antisense transcript |
US9758546B2 (en) | 2013-10-21 | 2017-09-12 | Ionis Pharmaceuticals, Inc. | Method for solution phase detritylation of oligomeric compounds |
EP3502270B1 (en) | 2013-10-21 | 2020-03-18 | The General Hospital Corporation | Methods relating to circulating tumor cell clusters and the treatment of cancer |
WO2015066708A1 (en) | 2013-11-04 | 2015-05-07 | Northwestern University | Quantification and spatio-temporal tracking of a target using a spherical nucleic acid (sna) |
CA2932122C (en) | 2013-12-03 | 2022-04-19 | Northwestern University | Liposomal particles, methods of making same and uses thereof |
CA2844640A1 (en) | 2013-12-06 | 2015-06-06 | The University Of British Columbia | Method for treatment of castration-resistant prostate cancer |
WO2015085183A2 (en) | 2013-12-06 | 2015-06-11 | Swift Biosciences, Inc. | Cleavable competitor polynucleotides |
AU2014362262B2 (en) | 2013-12-12 | 2021-05-13 | Alnylam Pharmaceuticals, Inc. | Complement component iRNA compositions and methods of use thereof |
CN106456694B (en) | 2013-12-20 | 2020-06-30 | 通用医疗公司 | Methods and assays relating to circulating tumor cells |
CN111394355A (en) | 2013-12-24 | 2020-07-10 | Ionis制药公司 | Modulation of angiopoietin-like 3 expression |
DK3102197T3 (en) | 2014-02-04 | 2018-11-19 | Genentech Inc | Smoothened mutant and methods for its use |
CN113057959A (en) | 2014-02-11 | 2021-07-02 | 阿尔尼拉姆医药品有限公司 | Ketohexokinase (KHK) iRNA compositions and methods of use thereof |
WO2015142910A1 (en) | 2014-03-17 | 2015-09-24 | Isis Pharmaceuticals, Inc. | Bicyclic carbocyclic nucleosides and oligomeric compounds prepared therefrom |
US10006027B2 (en) | 2014-03-19 | 2018-06-26 | Ionis Pharmaceuticals, Inc. | Methods for modulating Ataxin 2 expression |
CN106103717A (en) | 2014-03-19 | 2016-11-09 | Ionis制药公司 | For regulating the compositions that ataxin 2 is expressed |
SG11201608109TA (en) | 2014-04-01 | 2016-10-28 | Ionis Pharmaceuticals Inc | Compositions for modulating sod-1 expression |
EP3129493B1 (en) | 2014-04-09 | 2021-07-07 | The Scripps Research Institute | Import of unnatural or modified nucleoside triphosphates into cells via nucleic acid triphosphate transporters |
WO2015161170A2 (en) | 2014-04-17 | 2015-10-22 | Isis Pharmaceuticals, Inc. | Compositions and methods for modulation of smn2 splicing in a subject |
WO2015164693A1 (en) | 2014-04-24 | 2015-10-29 | Isis Pharmaceuticals, Inc. | Oligomeric compounds comprising alpha-beta-constrained nucleic acid |
HUE052709T2 (en) | 2014-05-01 | 2021-05-28 | Ionis Pharmaceuticals Inc | Conjugates of modified antisense oligonucleotides and their use for modulating pkk expression |
WO2015168514A1 (en) | 2014-05-01 | 2015-11-05 | Isis Pharmaceuticals, Inc. | Method for synthesis of reactive conjugate clusters |
KR102149571B1 (en) | 2014-05-01 | 2020-08-31 | 아이오니스 파마수티컬즈, 인코포레이티드 | Compositions and methods for modulating growth hormone receptor expression |
KR102369736B1 (en) | 2014-05-01 | 2022-03-02 | 아이오니스 파마수티컬즈, 인코포레이티드 | Compositions and methods for modulating complement factor b expression |
SI3137605T1 (en) | 2014-05-01 | 2021-02-26 | Ionis Pharmaceuticals, Inc. | Compositions and methods for modulating angiopoietin-like 3 expression |
WO2015175510A1 (en) | 2014-05-12 | 2015-11-19 | Alnylam Pharmaceuticals, Inc. | Methods and compositions for treating a serpinc1-associated disorder |
SG10202104570TA (en) | 2014-05-22 | 2021-06-29 | Alnylam Pharmaceuticals Inc | Angiotensinogen (agt) irna compositions and methods of use thereof |
CN106659758A (en) | 2014-06-02 | 2017-05-10 | 儿童医疗中心有限公司 | Methods and compositions for immunomodulation |
CN106535876B (en) | 2014-06-04 | 2020-09-11 | 埃克西奎雷股份有限公司 | Multivalent delivery of immunomodulators through liposomal spherical nucleic acids for prophylactic or therapeutic applications |
AU2015272128B2 (en) | 2014-06-10 | 2021-10-28 | Erasmus University Medical Center Rotterdam | Antisense oligonucleotides useful in treatment of Pompe disease |
TW201620526A (en) | 2014-06-17 | 2016-06-16 | 愛羅海德研究公司 | Compositions and methods for inhibiting gene expression of alpha-1 antitrypsin |
EP4159741A1 (en) | 2014-07-16 | 2023-04-05 | ModernaTX, Inc. | Method for producing a chimeric polynucleotide encoding a polypeptide having a triazole-containing internucleotide linkage |
US9951327B1 (en) | 2014-07-17 | 2018-04-24 | Integrated Dna Technologies, Inc. | Efficient and rapid method for assembling and cloning double-stranded DNA fragments |
US20170210788A1 (en) | 2014-07-23 | 2017-07-27 | Modernatx, Inc. | Modified polynucleotides for the production of intrabodies |
US10653747B2 (en) | 2014-07-31 | 2020-05-19 | Uab Research Foundation | ApoE mimetic peptides and higher potency to clear plasma cholesterol |
EP3183007B1 (en) | 2014-08-19 | 2020-06-17 | Northwestern University | Protein/oligonucleotide core-shell nanoparticle therapeutics |
EP3185910A4 (en) | 2014-08-29 | 2018-01-10 | Children's Medical Center Corporation | Methods and compositions for the treatment of cancer |
WO2016033424A1 (en) | 2014-08-29 | 2016-03-03 | Genzyme Corporation | Methods for the prevention and treatment of major adverse cardiovascular events using compounds that modulate apolipoprotein b |
KR102631505B1 (en) | 2014-08-29 | 2024-02-01 | 알닐람 파마슈티칼스 인코포레이티드 | Methods of treating transthyretin(ttr) mediated amyloidosis |
EP3191591A1 (en) | 2014-09-12 | 2017-07-19 | Alnylam Pharmaceuticals, Inc. | Polynucleotide agents targeting complement component c5 and methods of use thereof |
US10533172B2 (en) | 2014-09-18 | 2020-01-14 | The University Of British Columbia | Allele-specific therapy for huntington disease haplotypes |
JOP20200115A1 (en) | 2014-10-10 | 2017-06-16 | Alnylam Pharmaceuticals Inc | Compositions And Methods For Inhibition Of HAO1 (Hydroxyacid Oxidase 1 (Glycolate Oxidase)) Gene Expression |
WO2016061487A1 (en) | 2014-10-17 | 2016-04-21 | Alnylam Pharmaceuticals, Inc. | Polynucleotide agents targeting aminolevulinic acid synthase-1 (alas1) and uses thereof |
WO2016069694A2 (en) | 2014-10-30 | 2016-05-06 | Alnylam Pharmaceuticals, Inc. | Polynucleotide agents targeting serpinc1 (at3) and methods of use thereof |
US20170312297A1 (en) | 2014-11-10 | 2017-11-02 | Glaxosmithkline Intellectual Property (No. 2) Limited | Long Acting Pharmaceutical Compositions For Hepatitis C |
WO2016075584A1 (en) | 2014-11-10 | 2016-05-19 | Glaxosmithkline Intellectual Property (No.2) Limited | Combination long acting compositions and methods for hepatitis c |
JOP20200092A1 (en) | 2014-11-10 | 2017-06-16 | Alnylam Pharmaceuticals Inc | HEPATITIS B VIRUS (HBV) iRNA COMPOSITIONS AND METHODS OF USE THEREOF |
CA2968114A1 (en) | 2014-11-17 | 2016-05-26 | Alnylam Pharmaceuticals, Inc. | Apolipoprotein c3 (apoc3) irna compositions and methods of use thereof |
AU2015349680A1 (en) | 2014-11-21 | 2017-06-08 | Northwestern University | The sequence-specific cellular uptake of spherical nucleic acid nanoparticle conjugates |
WO2016086104A1 (en) | 2014-11-25 | 2016-06-02 | Ionis Pharmaceuticals, Inc. | Modulation of ube3a-ats expression |
WO2016100716A1 (en) | 2014-12-18 | 2016-06-23 | Vasant Jadhav | Reversirtm compounds |
US9688707B2 (en) | 2014-12-30 | 2017-06-27 | Ionis Pharmaceuticals, Inc. | Bicyclic morpholino compounds and oligomeric compounds prepared therefrom |
US10793855B2 (en) | 2015-01-06 | 2020-10-06 | Ionis Pharmaceuticals, Inc. | Compositions for modulating expression of C9ORF72 antisense transcript |
US10538763B2 (en) | 2015-01-16 | 2020-01-21 | Ionis Pharmaceuticals, Inc. | Compounds and methods for modulation of DUX4 |
EP3247988A4 (en) | 2015-01-23 | 2018-12-19 | Vanderbilt University | A robust interferometer and methods of using same |
AU2016219263B2 (en) | 2015-02-13 | 2022-12-01 | Alnylam Pharmaceuticals, Inc. | Patatin-like phospholipase domain containing 3 (PNPLA3) iRNA compositions and methods of use thereof |
US10450342B2 (en) | 2015-02-23 | 2019-10-22 | Ionis Pharmaceuticals, Inc. | Method for solution phase detritylation of oligomeric compounds |
MX2017011010A (en) | 2015-02-26 | 2017-10-20 | Ionis Pharmaceuticals Inc | Allele specific modulators of p23h rhodopsin. |
US11129844B2 (en) | 2015-03-03 | 2021-09-28 | Ionis Pharmaceuticals, Inc. | Compositions and methods for modulating MECP2 expression |
KR20180020125A (en) | 2015-03-27 | 2018-02-27 | 프레지던트 앤드 펠로우즈 오브 하바드 칼리지 | Modified T cells and methods for their manufacture and use |
WO2016164746A1 (en) | 2015-04-08 | 2016-10-13 | Alnylam Pharmaceuticals, Inc. | Compositions and methods for inhibiting expression of the lect2 gene |
WO2016167780A1 (en) | 2015-04-16 | 2016-10-20 | Ionis Pharmaceuticals, Inc. | Compositions for modulating expression of c9orf72 antisense transcript |
KR102258516B1 (en) | 2015-04-16 | 2021-05-31 | 아이오니스 파마수티컬즈, 인코포레이티드 | Compositions for modulating c9orf72 expression |
WO2016201301A1 (en) | 2015-06-12 | 2016-12-15 | Alnylam Pharmaceuticals, Inc. | Complement component c5 irna compositions and methods of use thereof |
EP3310918B1 (en) | 2015-06-18 | 2020-08-05 | Alnylam Pharmaceuticals, Inc. | Polynucleotide agents targeting hydroxyacid oxidase (glycolate oxidase, hao1) and methods of use thereof |
WO2016209862A1 (en) | 2015-06-23 | 2016-12-29 | Alnylam Pharmaceuticals, Inc. | Glucokinase (gck) irna compositions and methods of use thereof |
US10494632B2 (en) | 2015-07-10 | 2019-12-03 | Alnylam Pharmaceuticals, Inc. | Insulin-like growth factor binding protein, acid labile subunit (IGFALS) compositions and methods of use thereof |
JP2018520683A (en) | 2015-07-17 | 2018-08-02 | アルニラム ファーマスーティカルズ インコーポレイテッドAlnylam Pharmaceuticals, Inc. | Multi-target single-body conjugates |
US20180237774A1 (en) | 2015-08-04 | 2018-08-23 | Yeda Research And Development Co. Ltd. | Methods of screening for riboswitches and attenuators |
US10130651B2 (en) | 2015-08-07 | 2018-11-20 | Arrowhead Pharmaceuticals, Inc. | RNAi Therapy for Hepatitis B Virus Infection |
CN114525280A (en) | 2015-09-02 | 2022-05-24 | 阿尔尼拉姆医药品有限公司 | iRNA compositions of programmed cell death 1 ligand 1(PD-L1) and methods of use thereof |
EP4285912A2 (en) | 2015-09-25 | 2023-12-06 | Ionis Pharmaceuticals, Inc. | Compositions and methods for modulating ataxin 3 expression |
CN108271351B (en) | 2015-10-08 | 2021-10-26 | Ionis 制药公司 | Compounds and methods for modulating angiotensinogen expression |
EP3370734B1 (en) | 2015-11-05 | 2023-01-04 | Children's Hospital Los Angeles | Antisense oligo for use in treating acute myeloid leukemia |
JP2018531605A (en) | 2015-11-06 | 2018-11-01 | アイオーニス ファーマシューティカルズ, インコーポレーテッドIonis Pharmaceuticals,Inc. | Regulation of apolipoprotein (a) expression |
US11058709B1 (en) | 2015-12-04 | 2021-07-13 | Ionis Pharmaceuticals, Inc. | Methods of treating breast cancer |
CA3007152A1 (en) | 2015-12-07 | 2017-06-15 | Erasmus University Medical Center Rotterdam | Enzymatic replacement therapy and antisense therapy for pompe disease |
US11761007B2 (en) | 2015-12-18 | 2023-09-19 | The Scripps Research Institute | Production of unnatural nucleotides using a CRISPR/Cas9 system |
CA3006599A1 (en) | 2016-01-05 | 2017-07-13 | Ionis Pharmaceuticals, Inc. | Methods for reducing lrrk2 expression |
WO2017132483A1 (en) | 2016-01-29 | 2017-08-03 | Vanderbilt University | Free-solution response function interferometry |
JP2019509721A (en) | 2016-02-04 | 2019-04-11 | キュリス,インコーポレイテッド | Mutant smoothened and method of using the same |
WO2017161168A1 (en) | 2016-03-16 | 2017-09-21 | Ionis Pharmaceuticals, Inc. | Modulation of dyrk1b expression |
US10961271B2 (en) | 2016-03-16 | 2021-03-30 | Ionis Pharmaceuticals, Inc. | Methods of modulating KEAP1 |
MA45295A (en) | 2016-04-19 | 2019-02-27 | Alnylam Pharmaceuticals Inc | HIGH DENSITY LIPOPROTEIN BINDING PROTEIN (HDLBP / VIGILINE) RNA COMPOSITION AND METHODS FOR USING THEM |
EP3469083A1 (en) | 2016-06-10 | 2019-04-17 | Alnylam Pharmaceuticals, Inc. | COMPLEMENT COMPONENT C5 iRNA COMPOSITIONS AND METHODS OF USE THEREOF FOR TREATING PAROXYSMAL NOCTURNAL HEMOGLOBINURIA (PNH) |
EP3471781A4 (en) | 2016-06-17 | 2020-05-06 | Ionis Pharmaceuticals, Inc. | Modulation of gys1 expression |
ES2929047T3 (en) | 2016-06-24 | 2022-11-24 | Scripps Research Inst | Novel nucleoside triphosphate transporter and uses thereof |
WO2018015936A2 (en) | 2016-07-21 | 2018-01-25 | Maxcyte, Inc. | Methods and compositions for modifying genomic dna |
JOP20170161A1 (en) | 2016-08-04 | 2019-01-30 | Arrowhead Pharmaceuticals Inc | RNAi Agents for Hepatitis B Virus Infection |
NL2017295B1 (en) | 2016-08-05 | 2018-02-14 | Univ Erasmus Med Ct Rotterdam | Antisense oligomeric compound for Pompe disease |
NL2017294B1 (en) | 2016-08-05 | 2018-02-14 | Univ Erasmus Med Ct Rotterdam | Natural cryptic exon removal by pairs of antisense oligonucleotides. |
WO2018039629A2 (en) | 2016-08-25 | 2018-03-01 | Northwestern University | Micellar spherical nucleic acids from thermoresponsive, traceless templates |
WO2018055577A1 (en) | 2016-09-23 | 2018-03-29 | Synthena Ag | Mixed tricyclo-dna, 2'-modified rna oligonucleotide compositions and uses thereof |
EP3522898A4 (en) | 2016-10-06 | 2020-05-27 | Ionis Pharmaceuticals, Inc. | Method of conjugating oligomeric compounds |
JOP20190104A1 (en) | 2016-11-10 | 2019-05-07 | Ionis Pharmaceuticals Inc | Compounds and methods for reducing atxn3 expression |
TWI788312B (en) | 2016-11-23 | 2023-01-01 | 美商阿尼拉製藥公司 | SERPINA1 iRNA COMPOSITIONS AND METHODS OF USE THEREOF |
EP3548620A4 (en) | 2016-12-02 | 2020-07-22 | Cold Spring Harbor Laboratory | Modulation of lnc05 expression |
KR20230166146A (en) | 2016-12-16 | 2023-12-06 | 알닐람 파마슈티칼스 인코포레이티드 | Methods for treating or preventing ttr-associated diseases using transthyretin(ttr) irna compositions |
KR20190098748A (en) | 2017-01-10 | 2019-08-22 | 애로우헤드 파마슈티컬스 인코포레이티드 | Alpha-1 antitrypsin (AAT) RNAi agent, compositions comprising AAT RNAi agent, and methods of use |
EP3571321A1 (en) | 2017-01-23 | 2019-11-27 | Regeneron Pharmaceuticals, Inc. | Hydroxysteroid 17-beta dehydrogenase 13 (hsd17b13) variants and uses thereof |
WO2018165564A1 (en) | 2017-03-09 | 2018-09-13 | Ionis Pharmaceuticals, Inc. | Morpholino modified oligomeric compounds |
WO2018183969A1 (en) | 2017-03-30 | 2018-10-04 | California Institute Of Technology | Barcoded rapid assay platform for efficient analysis of candidate molecules and methods of making and using the platform |
CN110913898B (en) | 2017-04-18 | 2024-04-05 | 阿尔尼拉姆医药品有限公司 | Methods of treating subjects having Hepatitis B Virus (HBV) infection |
WO2018193428A1 (en) | 2017-04-20 | 2018-10-25 | Synthena Ag | Modified oligomeric compounds comprising tricyclo-dna nucleosides and uses thereof |
EP3612546B1 (en) | 2017-04-20 | 2022-07-13 | Synthena AG | Modified oligomeric compounds comprising tricyclo-dna nucleosides and uses thereof |
US20200131555A1 (en) | 2017-07-11 | 2020-04-30 | Synthorx, Inc. | Incorporation of unnatural nucleotides and methods thereof |
MX2020000387A (en) | 2017-07-13 | 2020-08-17 | Univ Northwestern | General and direct method for preparing oligonucleotide-functiona lized metal-organic framework nanoparticles. |
EP3652317A1 (en) | 2017-07-13 | 2020-05-20 | Alnylam Pharmaceuticals, Inc. | Lactate dehydrogenase a (ldha) irna compositions and methods of use thereof |
TWI829642B (en) * | 2017-07-24 | 2024-01-21 | 美商寬騰矽公司 | High intensity labeled reactant compositions and methods for sequencing |
SG11202000939PA (en) | 2017-08-03 | 2020-02-27 | Synthorx Inc | Cytokine conjugates for the treatment of proliferative and infectious diseases |
WO2019036613A1 (en) | 2017-08-18 | 2019-02-21 | Ionis Pharmaceuticals, Inc. | Modulation of the notch signaling pathway for treatment of respiratory disorders |
US10517889B2 (en) | 2017-09-08 | 2019-12-31 | Ionis Pharmaceuticals, Inc. | Modulators of SMAD7 expression |
WO2019060442A1 (en) | 2017-09-19 | 2019-03-28 | Alnylam Pharmaceuticals, Inc. | Compositions and methods for treating transthyretin (ttr) mediated amyloidosis |
AU2018360697A1 (en) | 2017-11-01 | 2020-05-14 | Alnylam Pharmaceuticals, Inc. | Complement component C3 iRNA compositions and methods of use thereof |
TWI809004B (en) | 2017-11-09 | 2023-07-21 | 美商Ionis製藥公司 | Compounds and methods for reducing snca expression |
WO2019099610A1 (en) | 2017-11-16 | 2019-05-23 | Alnylam Pharmaceuticals, Inc. | Kisspeptin 1 (kiss1) irna compositions and methods of use thereof |
WO2019100039A1 (en) | 2017-11-20 | 2019-05-23 | Alnylam Pharmaceuticals, Inc. | Serum amyloid p component (apcs) irna compositions and methods of use thereof |
MX2020006012A (en) | 2017-12-18 | 2020-09-14 | Alnylam Pharmaceuticals Inc | High mobility group box-1 (hmgb1) irna compositions and methods of use thereof. |
WO2019126641A2 (en) | 2017-12-21 | 2019-06-27 | Ionis Pharmaceuticals, Inc. | Modulation of frataxin expression |
AU2019206731A1 (en) | 2018-01-15 | 2020-07-30 | Ionis Pharmaceuticals, Inc. | Modulators of DNM2 expression |
US20190233816A1 (en) | 2018-01-26 | 2019-08-01 | Massachusetts Institute Of Technology | Structure-guided chemical modification of guide rna and its applications |
SG11202007728QA (en) | 2018-02-26 | 2020-09-29 | Synthorx Inc | Il-15 conjugates and uses thereof |
TW202000199A (en) | 2018-03-02 | 2020-01-01 | 美商Ionis製藥公司 | Modulators of IRF4 expression |
EP3759127A4 (en) | 2018-03-02 | 2022-03-30 | Ionis Pharmaceuticals, Inc. | Compounds and methods for the modulation of amyloid-beta precursor protein |
WO2019183440A1 (en) | 2018-03-22 | 2019-09-26 | Ionis Pharmaceuticals, Inc. | Methods for modulating fmr1 expression |
JP2021520781A (en) | 2018-04-06 | 2021-08-26 | チルドレンズ メディカル センター コーポレーションChildren’S Medical Center Corporation | Compositions and Methods for Somatic Cell Reprogramming and Imprinting Modulation |
CA3094020A1 (en) | 2018-04-11 | 2019-10-17 | Ionis Pharmaceuticals, Inc. | Modulators of ezh2 expression |
TW202016301A (en) | 2018-05-07 | 2020-05-01 | 美商阿里拉姆製藥股份有限公司 | Extrahepatic delivery |
BR112020020957B1 (en) | 2018-05-09 | 2022-05-10 | Ionis Pharmaceuticals, Inc | Oligomeric compounds, population and pharmaceutical composition thereof and their uses |
AU2019266307A1 (en) | 2018-05-09 | 2020-11-19 | Ionis Pharmaceuticals, Inc. | Compounds and methods for reducing ATXN3 expression |
TW202016304A (en) | 2018-05-14 | 2020-05-01 | 美商阿尼拉製藥公司 | Angiotensinogen (agt) irna compositions and methods of use thereof |
AU2019287635A1 (en) | 2018-06-14 | 2020-12-17 | Ionis Pharmaceuticals, Inc. | Compounds and methods for increasing STMN2 expression |
SG11202011864XA (en) | 2018-06-27 | 2020-12-30 | Ionis Pharmaceuticals Inc | Compounds and methods for reducing lrrk2 expression |
WO2020023737A1 (en) | 2018-07-25 | 2020-01-30 | Ionis Pharmaceuticals, Inc. | Compounds and methods for reducing atxn2 expression |
TW202023574A (en) | 2018-08-13 | 2020-07-01 | 美商阿尼拉製藥公司 | Hepatitis b virus (hbv) dsrna agent compositions and methods of use thereof |
WO2020037125A1 (en) | 2018-08-16 | 2020-02-20 | Alnylam Pharmaceuticals Inc. | Compositions and methods for inhibiting expression of the lect2 gene |
SG11202102531WA (en) | 2018-09-14 | 2021-04-29 | Univ Northwestern | Programming protein polymerization with dna |
AU2019344776A1 (en) | 2018-09-18 | 2021-01-21 | Alnylam Pharmaceuticals, Inc. | Ketohexokinase (KHK) iRNA compositions and methods of use thereof |
TW202028465A (en) | 2018-09-28 | 2020-08-01 | 美商阿尼拉製藥公司 | Transthyretin (ttr) irna compositions and methods of use thereof for treating or preventing ttr-associated ocular diseases |
US10913951B2 (en) | 2018-10-31 | 2021-02-09 | University of Pittsburgh—of the Commonwealth System of Higher Education | Silencing of HNF4A-P2 isoforms with siRNA to improve hepatocyte function in liver failure |
TW202028222A (en) | 2018-11-14 | 2020-08-01 | 美商Ionis製藥公司 | Modulators of foxp3 expression |
TW202039840A (en) | 2018-11-15 | 2020-11-01 | 美商伊奧尼斯醫藥公司 | Modulators of irf5 expression |
US20210332495A1 (en) | 2018-12-06 | 2021-10-28 | Northwestern University | Protein Crystal Engineering Through DNA Hybridization Interactions |
SI3897672T1 (en) | 2018-12-20 | 2024-02-29 | Humabs Biomed Sa | Combination hbv therapy |
EP3898977A1 (en) | 2018-12-20 | 2021-10-27 | Praxis Precision Medicines, Inc. | Compositions and methods for the treatment of kcnt1 related disorders |
MX2021008628A (en) | 2019-01-16 | 2021-11-17 | Genzyme Corp | Serpinc1 irna compositions and methods of use thereof. |
JP2022518056A (en) | 2019-01-23 | 2022-03-11 | クアンタム-エスアイ インコーポレイテッド | Reaction Compositions and Methods for High Intensity Labeled Sequencing |
CA3128093A1 (en) | 2019-01-31 | 2020-08-06 | Ionis Pharmaceuticals, Inc. | Modulators of yap1 expression |
KR20210123299A (en) | 2019-02-06 | 2021-10-13 | 신톡스, 인크. | IL-2 conjugates and methods of use thereof |
SG11202108450SA (en) | 2019-02-27 | 2021-09-29 | Ionis Pharmaceuticals Inc | Modulators of malat1 expression |
US20220228140A1 (en) | 2019-03-29 | 2022-07-21 | Mitsubishi Tanabe Pharma Corporation | Compound, method and pharmaceutical composition for modulating expression of dux4 |
CN117431244A (en) | 2019-03-29 | 2024-01-23 | Ionis制药公司 | Compounds and methods for modulating UBE3A-ATS |
KR20220036914A (en) | 2019-05-13 | 2022-03-23 | 비르 바이오테크놀로지, 인코포레이티드 | Compositions and methods for treating hepatitis B virus (HBV) infection |
CA3138915A1 (en) | 2019-05-17 | 2020-11-26 | Alnylam Pharmaceuticals, Inc. | Oral delivery of oligonucleotides |
BR112021025130A2 (en) | 2019-06-14 | 2022-03-15 | Scripps Research Inst | Reagents and methods for replication, transcription and translation in semisynthetic organisms |
US20220305107A1 (en) | 2019-06-18 | 2022-09-29 | Janssen Sciences Ireland Unlimited Company | COMBINATION OF HEPATITIS B VIRUS (HBV) VACCINES AND HBV-TARGETING RNAi |
EP3986562A1 (en) | 2019-06-18 | 2022-04-27 | Janssen Sciences Ireland Unlimited Company | Combination of hepatitis b virus (hbv) vaccines and hbv-targeting rnai |
WO2021021673A1 (en) | 2019-07-26 | 2021-02-04 | Ionis Pharmaceuticals, Inc. | Compounds and methods for modulating gfap |
WO2021022109A1 (en) | 2019-08-01 | 2021-02-04 | Alnylam Pharmaceuticals, Inc. | SERPIN FAMILY F MEMBER 2 (SERPINF2) iRNA COMPOSITIONS AND METHODS OF USE THEREOF |
WO2021022108A2 (en) | 2019-08-01 | 2021-02-04 | Alnylam Pharmaceuticals, Inc. | CARBOXYPEPTIDASE B2 (CPB2) iRNA COMPOSITIONS AND METHODS OF USE THEREOF |
WO2021030522A1 (en) | 2019-08-13 | 2021-02-18 | Alnylam Pharmaceuticals, Inc. | SMALL RIBOSOMAL PROTEIN SUBUNIT 25 (RPS25) iRNA AGENT COMPOSITIONS AND METHODS OF USE THEREOF |
KR20220047598A (en) | 2019-08-15 | 2022-04-18 | 신톡스, 인크. | Immuno-oncology Combination Therapy Using IL-2 Conjugates |
JP2022544587A (en) | 2019-08-15 | 2022-10-19 | アイオーニス ファーマシューティカルズ, インコーポレーテッド | Bond-modified oligomeric compounds and uses thereof |
US20210054040A1 (en) | 2019-08-23 | 2021-02-25 | Synthorx, Inc. | Novel il-15 conjugates and uses thereof |
WO2021046122A1 (en) | 2019-09-03 | 2021-03-11 | Alnylam Pharmaceuticals, Inc. | Compositions and methods for inhibiting expression of the lect2 gene |
KR20220061158A (en) | 2019-09-10 | 2022-05-12 | 신톡스, 인크. | IL-2 conjugates and methods of use for treating autoimmune diseases |
WO2021067747A1 (en) | 2019-10-04 | 2021-04-08 | Alnylam Pharmaceuticals, Inc. | Compositions and methods for silencing ugt1a1 gene expression |
EP4045652A1 (en) | 2019-10-18 | 2022-08-24 | Alnylam Pharmaceuticals, Inc. | Solute carrier family member irna compositions and methods of use thereof |
BR112022007540A2 (en) | 2019-10-22 | 2022-07-12 | Alnylam Pharmaceuticals Inc | COMPONENTS COMPLEMENTARY C3 IRNA COMPOSITIONS AND METHODS OF USE THEREOF |
CA3155921A1 (en) | 2019-11-01 | 2021-05-06 | Alnylam Pharmaceuticals, Inc. | Huntingtin (htt) irna agent compositions and methods of use thereof |
EP4051796A1 (en) | 2019-11-01 | 2022-09-07 | Alnylam Pharmaceuticals, Inc. | Compositions and methods for silencing dnajb1-prkaca fusion gene expression |
JP2022554272A (en) | 2019-11-04 | 2022-12-28 | シンソークス, インコーポレイテッド | Interleukin 10 conjugates and uses thereof |
WO2021092145A1 (en) | 2019-11-06 | 2021-05-14 | Alnylam Pharmaceuticals, Inc. | Transthyretin (ttr) irna composition and methods of use thereof for treating or preventing ttr-associated ocular diseases |
KR20220110749A (en) | 2019-11-06 | 2022-08-09 | 알닐람 파마슈티칼스 인코포레이티드 | extrahepatic transmission |
EP4061945A1 (en) | 2019-11-22 | 2022-09-28 | Alnylam Pharmaceuticals, Inc. | Ataxin3 (atxn3) rnai agent compositions and methods of use thereof |
JP2023506181A (en) | 2019-12-13 | 2023-02-15 | アルナイラム ファーマシューティカルズ, インコーポレイテッド | Human chromosome 9 open reading frame 72 (C9ORF72) iRNA agent compositions and methods of use thereof |
WO2021126734A1 (en) | 2019-12-16 | 2021-06-24 | Alnylam Pharmaceuticals, Inc. | Patatin-like phospholipase domain containing 3 (pnpla3) irna compositions and methods of use thereof |
US20230057461A1 (en) | 2020-01-27 | 2023-02-23 | The U.S.A., As Represented By The Secretary, Department Of Health And Human Services | Rab13 and net1 antisense oligonucleotides to treat metastatic cancer |
WO2021154941A1 (en) | 2020-01-31 | 2021-08-05 | Alnylam Pharmaceuticals, Inc. | Complement component c5 irna compositions for use in the treatment of amyotrophic lateral sclerosis (als) |
EP4103714A1 (en) | 2020-02-10 | 2022-12-21 | Alnylam Pharmaceuticals, Inc. | Compositions and methods for silencing vegf-a expression |
EP4107265A1 (en) | 2020-02-18 | 2022-12-28 | Alnylam Pharmaceuticals, Inc. | Apolipoprotein c3 (apoc3) irna compositions and methods of use thereof |
CR20220485A (en) | 2020-02-28 | 2022-11-10 | Ionis Pharmaceuticals Inc | Compounds and methods for modulating smn2 |
EP4114947A1 (en) | 2020-03-05 | 2023-01-11 | Alnylam Pharmaceuticals, Inc. | Complement component c3 irna compositions and methods of use thereof for treating or preventing complement component c3-associated diseases |
EP4114948A1 (en) | 2020-03-06 | 2023-01-11 | Alnylam Pharmaceuticals, Inc. | Ketohexokinase (khk) irna compositions and methods of use thereof |
EP4121534A1 (en) | 2020-03-18 | 2023-01-25 | Alnylam Pharmaceuticals, Inc. | Compositions and methods for treating subjects having a heterozygous alanine-glyoxylate aminotransferase gene (agxt) variant |
JP2023519274A (en) | 2020-03-26 | 2023-05-10 | アルナイラム ファーマシューティカルズ, インコーポレイテッド | CORONAVIRUS iRNA COMPOSITIONS AND METHODS OF USE THEREOF |
WO2021202443A2 (en) | 2020-03-30 | 2021-10-07 | Alnylam Pharmaceucticals, Inc. | Compositions and methods for silencing dnajc15 gene expression |
JP2023520582A (en) | 2020-04-06 | 2023-05-17 | アルナイラム ファーマシューティカルズ, インコーポレイテッド | Compositions and methods for silencing MYOC expression |
US20230159933A1 (en) | 2020-04-07 | 2023-05-25 | Alnylam Pharmaceuticals, Inc. | Compositions and methods for silencing scn9a expression |
EP4133077A1 (en) | 2020-04-07 | 2023-02-15 | Alnylam Pharmaceuticals, Inc. | Transmembrane serine protease 2 (tmprss2) irna compositions and methods of use thereof |
EP4133076A1 (en) | 2020-04-07 | 2023-02-15 | Alnylam Pharmaceuticals, Inc. | Angiotensin-converting enzyme 2 (ace2) irna compositions and methods of use thereof |
CN115955972A (en) | 2020-04-27 | 2023-04-11 | 阿尔尼拉姆医药品有限公司 | Apolipoprotein E (APOE) iRNA agent compositions and methods of use thereof |
AU2021265813A1 (en) | 2020-04-30 | 2022-11-10 | Alnylam Pharmaceuticals, Inc. | Complement factor B (CFB) iRNA compositions and methods of use thereof |
CA3181546A1 (en) | 2020-05-01 | 2021-11-04 | Ionis Pharmaceuticals, Inc. | Compounds and methods for modulating atxn1 |
US20230227824A1 (en) | 2020-05-12 | 2023-07-20 | Mitsubishi Tanabe Pharma Corporation | Compound, method and pharmaceutical composition for regulating expression of ataxin 3 |
WO2021231691A1 (en) | 2020-05-15 | 2021-11-18 | Korro Bio, Inc. | Methods and compositions for the adar-mediated editing of retinoschisin 1 (rsi) |
WO2021231680A1 (en) | 2020-05-15 | 2021-11-18 | Korro Bio, Inc. | Methods and compositions for the adar-mediated editing of methyl-cpg binding protein 2 (mecp2) |
EP4150077A1 (en) | 2020-05-15 | 2023-03-22 | Korro Bio, Inc. | Methods and compositions for the adar-mediated editing of transmembrane channel-like protein 1 (tmc1) |
WO2021231679A1 (en) | 2020-05-15 | 2021-11-18 | Korro Bio, Inc. | Methods and compositions for the adar-mediated editing of gap junction protein beta 2 (gjb2) |
EP4150078A1 (en) | 2020-05-15 | 2023-03-22 | Korro Bio, Inc. | Methods and compositions for the adar-mediated editing of argininosuccinate lyase (asl) |
WO2021231692A1 (en) | 2020-05-15 | 2021-11-18 | Korro Bio, Inc. | Methods and compositions for the adar-mediated editing of otoferlin (otof) |
CA3162416C (en) | 2020-05-15 | 2023-07-04 | Korro Bio, Inc. | Methods and compositions for the adar-mediated editing of argininosuccinate synthetase (ass1) |
EP4150086A1 (en) | 2020-05-15 | 2023-03-22 | Korro Bio, Inc. | Methods and compositions for the adar-mediated editing of leucine rich repeat kinase 2 (lrrk2) |
US20230183707A1 (en) | 2020-05-21 | 2023-06-15 | Alnylam Pharmaceuticals, Inc. | Compositions and methods for inhibiting marc1 gene expression |
AR122534A1 (en) | 2020-06-03 | 2022-09-21 | Triplet Therapeutics Inc | METHODS FOR THE TREATMENT OF NUCLEOTIDE REPEAT EXPANSION DISORDERS ASSOCIATED WITH MSH3 ACTIVITY |
JP2023530234A (en) | 2020-06-05 | 2023-07-14 | ザ・ブロード・インスティテュート・インコーポレイテッド | Compositions and methods for treating neoplasms |
EP4162050A1 (en) | 2020-06-09 | 2023-04-12 | Alnylam Pharmaceuticals, Inc. | Rnai compositions and methods of use thereof for delivery by inhalation |
CA3184289A1 (en) | 2020-06-18 | 2021-12-23 | Alnylam Pharmaceuticals, Inc. | Xanthine dehydrogenase (xdh) irna compositions and methods of use thereof |
KR20230042023A (en) | 2020-06-24 | 2023-03-27 | 비르 바이오테크놀로지, 인코포레이티드 | Engineered hepatitis B virus neutralizing antibodies and uses thereof |
AU2021296622A1 (en) | 2020-06-25 | 2023-02-23 | Synthorx, Inc. | Immuno oncology combination therapy with IL-2 conjugates and anti-EGFR antibodies |
WO2022006134A2 (en) | 2020-06-29 | 2022-01-06 | Ionis Pharmaceuticals, Inc. | Compounds and methods for modulating plp1 |
WO2022011214A1 (en) | 2020-07-10 | 2022-01-13 | Alnylam Pharmaceuticals, Inc. | Circular sirnas |
EP4217489A1 (en) | 2020-09-24 | 2023-08-02 | Alnylam Pharmaceuticals, Inc. | Dipeptidyl peptidase 4 (dpp4) irna compositions and methods of use thereof |
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WO2022147223A2 (en) | 2020-12-31 | 2022-07-07 | Alnylam Pharmaceuticals, Inc. | 2'-modified nucleoside based oligonucleotide prodrugs |
WO2022147214A2 (en) | 2020-12-31 | 2022-07-07 | Alnylam Pharmaceuticals, Inc. | Cyclic-disulfide modified phosphate based oligonucleotide prodrugs |
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WO2022174101A1 (en) | 2021-02-12 | 2022-08-18 | Synthorx, Inc. | Skin cancer combination therapy with il-2 conjugates and cemiplimab |
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AU2022249318A1 (en) | 2021-03-31 | 2023-10-12 | Entrada Therapeutics, Inc. | Cyclic cell penetrating peptides |
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US20230194709A9 (en) | 2021-06-29 | 2023-06-22 | Seagate Technology Llc | Range information detection using coherent pulse sets with selected waveform characteristics |
WO2023278410A1 (en) | 2021-06-29 | 2023-01-05 | Korro Bio, Inc. | Methods and compositions for adar-mediated editing |
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Family Cites Families (12)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4483964A (en) * | 1983-06-20 | 1984-11-20 | Chiron Corporation | Reactor system and method for polynucleotide synthesis |
US4517338A (en) * | 1983-06-20 | 1985-05-14 | Chiron Corporation | Multiple reactor system and method for polynucleotide synthesis |
US5430136A (en) * | 1984-10-16 | 1995-07-04 | Chiron Corporation | Oligonucleotides having selectably cleavable and/or abasic sites |
US5118605A (en) * | 1984-10-16 | 1992-06-02 | Chiron Corporation | Polynucleotide determination with selectable cleavage sites |
US5367066A (en) * | 1984-10-16 | 1994-11-22 | Chiron Corporation | Oligonucleotides with selectably cleavable and/or abasic sites |
US4775619A (en) * | 1984-10-16 | 1988-10-04 | Chiron Corporation | Polynucleotide determination with selectable cleavage sites |
US4910300A (en) * | 1985-12-11 | 1990-03-20 | Chiron Corporation | Method for making nucleic acid probes |
US5256549A (en) * | 1986-03-28 | 1993-10-26 | Chiron Corporation | Purification of synthetic oligomers |
US5359100A (en) * | 1987-10-15 | 1994-10-25 | Chiron Corporation | Bifunctional blocked phosphoramidites useful in making nucleic acid mutimers |
WO1995006659A1 (en) * | 1992-07-01 | 1995-03-09 | Isis Pharmaceuticals, Inc. | Amine-derivatized nucleosides and oligonucleosides |
NL9000156A (en) * | 1990-01-22 | 1991-08-16 | Rijksuniversiteit | Amino-protecting gps., esp. for nucleic acid synthesis - comprise ortho-tri:organo-silyl:oxy-methyl-aryl-carbonyl gps. |
US5430138A (en) * | 1990-07-27 | 1995-07-04 | Chiron Corporation | Hydroxyl-protecting groups attached to cytidine nucleotide compounds which are orthogonally removable |
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1995
- 1995-04-26 US US08/429,197 patent/US5594117A/en not_active Expired - Fee Related
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- 1995-08-24 WO PCT/US1995/010776 patent/WO1996006104A1/en active IP Right Grant
- 1995-08-24 CA CA002196806A patent/CA2196806A1/en not_active Abandoned
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US5594117A (en) | 1997-01-14 |
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EP0777674B1 (en) | 2001-11-28 |
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JP2006068022A (en) | 2006-03-16 |
US5597909A (en) | 1997-01-28 |
DE69524232D1 (en) | 2002-01-10 |
EP0777674A1 (en) | 1997-06-11 |
MX9701293A (en) | 1997-05-31 |
WO1996006104A1 (en) | 1996-02-29 |
ATE209655T1 (en) | 2001-12-15 |
JPH10504719A (en) | 1998-05-12 |
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