Compositions and methods for modulating SMN gene family expression
원문보기
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
C12N-015/113
A61K-031/7125
A61K-031/7115
A61K-031/712
A61K-031/7088
C07H-021/00
출원번호
US-0401194
(2013-05-16)
등록번호
US-10059941
(2018-08-28)
국제출원번호
PCT/US2013/041440
(2013-05-16)
국제공개번호
WO2013/173638
(2013-11-21)
발명자
/ 주소
Krieg, Arthur M.
Subramanian, Romesh
McSwiggen, James
Lee, Jeannie T.
출원인 / 주소
Translate Bio MA, Inc.
대리인 / 주소
Wolf, Greenfield & Sacks, P.C.
인용정보
피인용 횟수 :
0인용 특허 :
74
초록▼
Aspects of the invention provide single stranded oligonucleotides for activating or enhancing expression of SMN1 or SMN2. Further aspects provide compositions and kits comprising single stranded oligonucleotides for activating or enhancing expression of SMN1 or SMN2 that comprises exon 7. Methods fo
Aspects of the invention provide single stranded oligonucleotides for activating or enhancing expression of SMN1 or SMN2. Further aspects provide compositions and kits comprising single stranded oligonucleotides for activating or enhancing expression of SMN1 or SMN2 that comprises exon 7. Methods for modulating expression of SMN1 or SMN2 using the single stranded oligonucleotides are also provided. Further aspects of the invention provide methods for selecting a candidate oligonucleotide for activating or enhancing expression of SMN1 or SMN2.
대표청구항▼
1. A composition comprising a cell and a single stranded oligonucleotide, wherein the single stranded oligonucleotide is produced by a process comprising: synthesizing a single stranded oligonucleotide that:(a) has a sequence 5′-X-Y-Z, wherein X is any nucleotide, Y is a nucleotide sequence of 6 nuc
1. A composition comprising a cell and a single stranded oligonucleotide, wherein the single stranded oligonucleotide is produced by a process comprising: synthesizing a single stranded oligonucleotide that:(a) has a sequence 5′-X-Y-Z, wherein X is any nucleotide, Y is a nucleotide sequence of 6 nucleotides in length that is not a seed sequence of a human microRNA, and Z is a nucleotide sequence of 1 to 8 nucleotides in length,(b) is 100% complementary with a PRC2-associated region of an SMN gene, wherein the PRC2-associated region is a region of the SMN gene that has a sequence that occurs at a higher frequency in a sequencing reaction of products of an RNA-immunoprecipitation assay that employs an antibody that targets Ezh2 to immunoprecipitate RNA-associated PRC2 complexes from cells comprising the SMN gene compared to a control sequencing reaction of products of a control RNA-immunoprecipitation assay that employs a control antibody, and(c) is 8 to 15 nucleotides in length,wherein, during the synthesis, at least one nucleotide incorporated into the oligonucleotide is a nucleotide analogue and/or a modified internucleotide linkage is incorporated between at least two nucleotides. 2. The composition of claim 1, wherein the oligonucleotide does not comprise three or more consecutive guanosine nucleotides. 3. The composition of claim 1, wherein the oligonucleotide does not comprise four or more consecutive guanosine nucleotides. 4. The composition of claim 1, wherein the oligonucleotide is 8 to 10 nucleotides in length and all but 1, 2, or 3 of the nucleotides of the complementary sequence of the PRC2-associated region are cytosine or guanosine nucleotides. 5. The composition of claim 1, wherein at least one nucleotide of the oligonucleotide is a nucleotide analogue. 6. The composition of claim 5, wherein the at least one nucleotide analogue results in an increase in Tm of the oligonucleotide in a range of 1 to 5° C. compared with an oligonucleotide that does not have the at least one nucleotide analogue. 7. The composition of claim 1, wherein at least one nucleotide of the oligonucleotide comprises a 2′ O-methyl. 8. The composition of claim 1, wherein each nucleotide of the oligonucleotide comprises a 2′ O-methyl. 9. The composition of claim 1, wherein the oligonucleotide comprises at least one ribonucleotide, at least one deoxyribonucleotide, or at least one bridged nucleotide. 10. The composition of claim 9, wherein the bridged nucleotide is a LNA nucleotide, a cEt nucleotide or a ENA modified nucleotide. 11. The composition of claim 1, wherein each nucleotide of the oligonucleotide is a LNA nucleotide. 12. The composition of claim 1, wherein the nucleotides of the oligonucleotide comprise alternating deoxyribonucleotides and 2′-fluoro-deoxyribonucleotides. 13. The composition of claim 1, wherein the nucleotides of the oligonucleotide comprise alternating deoxyribonucleotides and 2′-O-methyl nucleotides. 14. The composition of claim 1, wherein the nucleotides of the oligonucleotide comprise alternating deoxyribonucleotides and ENA nucleotide analogues. 15. The composition of claim 1, wherein the nucleotides of the oligonucleotide comprise alternating deoxyribonucleotides and LNA nucleotides. 16. The composition of claim 12, wherein the 5′ nucleotide of the oligonucleotide is a deoxyribonucleotide. 17. The composition of claim 1, wherein the nucleotides of the oligonucleotide comprise alternating LNA nucleotides and 2′-O-methyl nucleotides. 18. The composition of claim 17, wherein the 5′ nucleotide of the oligonucleotide is a LNA nucleotide. 19. The composition of claim 1, wherein the nucleotides of the oligonucleotide comprise deoxyribonucleotides flanked by at least one LNA nucleotide on each of the 5′ and 3′ ends of the deoxyribonucleotides. 20. The composition of claim 1, further comprising phosphorothioate internucleotide linkages between at least two nucleotides. 21. The composition of claim 20, further comprising phosphorothioate internucleotide linkages between all nucleotides. 22. The composition of claim 1, wherein the nucleotide at the 3′ position of the oligonucleotide has a 3′ hydroxyl group. 23. The composition of claim 1, wherein the nucleotide at the 3′ position of the oligonucleotide has a 3′ thiophosphate. 24. The composition of claim 1, further comprising a biotin moiety conjugated to the 5′ nucleotide. 25. A composition comprising a cell and a single stranded oligonucleotide, wherein the single stranded oligonucleotide is produced by a process comprising: synthesizing a single stranded oligonucleotide that:(a) is 100% complementary with a PRC2-associated region of an SMN gene, wherein the PRC2-associated region is a region of the SMN gene that has a sequence that occurs at a higher frequency in a sequencing reaction of products of an RNA-immunoprecipitation assay that employs an antibody that targets Ezh2 to immunoprecipitate RNA-associated PRC2 complexes from cells comprising the SMN gene compared to a control sequencing reaction of products of a control RNA-immunoprecipitation assay that employs a control antibody,(b) is 8 to 15 nucleotides in length and has at least one of the following features i)-iv): i) a sequence that is 5′X-Y-Z, wherein X is any nucleotide and wherein X is anchored at the 5′ end of the oligonucleotide, Y is a nucleotide sequence of 6 nucleotides in length that is not a human seed sequence of a microRNA, and Z is a nucleotide sequence of 1 to 23 nucleotides in length;ii) a sequence that does not comprise three or more consecutive guanosine nucleotides;iii) a sequence that is complementary to a PRC2-associated region that encodes an RNA that forms a secondary structure comprising at least two single stranded loops; and/oriv) a sequence that has greater than 60% G-C content;wherein, during the synthesis, at least one nucleotide incorporated into the oligonucleotide is a nucleotide analogue and/or a modified internucleotide linkage is incorporated between at least two nucleotides. 26. The composition of claim 25, wherein the oligonucleotide has the sequence 5′X-Y-Z and at least one of features b), c) and d). 27. A composition comprising a cell and a single stranded oligonucleotide conjugated to a carrier, wherein the single stranded oligonucleotide is produced by a process comprising: synthesizing a single stranded oligonucleotide that:(a) has a sequence 5′-X-Y-Z, wherein X is any nucleotide, Y is a nucleotide sequence of 6 nucleotides in length that is not a seed sequence of a human microRNA, and Z is a nucleotide sequence of 1 to 8 nucleotides in length,(b) is 100% complementary with a PRC2-associated region of a human SMN gene, wherein the PRC2-associated region is a region of the SMN gene that has a sequence that occurs at a higher frequency in a sequencing reaction of products of an RNA-immunoprecipitation assay that employs an antibody that targets Ezh2 to immunoprecipitate RNA-associated PRC2 complexes from cells comprising the SMN gene compared to a control sequencing reaction of products of a control RNA-immunoprecipitation assay that employs a control antibody, and(c) is 8 to 15 nucleotides in length,wherein, during the synthesis, at least one nucleotide incorporated into the oligonucleotide is a nucleotide analogue and/or a modified internucleotide linkage is incorporated between at least two nucleotides. 28. The composition of claim 27, wherein the carrier is a peptide. 29. The composition of claim 27, wherein the carrier is a steroid. 30. A pharmaceutical composition comprising a composition of claim 27 and a pharmaceutically acceptable carrier. 31. A kit comprising a container housing the composition of claim 27. 32. A method of increasing expression of SMN1 or SMN2 messenger RNA (mRNA) in a human cell, the method comprising: delivering a single stranded oligonucleotide into the cell, wherein the oligonucleotide does not induce substantial cleavage or degradation of the SMN1 or SMN2 mRNA in the cell and wherein the single stranded oligonucleotide is produced by a process comprising:synthesizing a single stranded oligonucleotide that: (a) has a sequence 5′-X-Y-Z, wherein X is any nucleotide, Y is a nucleotide sequence of 6 nucleotides in length that is not a seed sequence of a human microRNA, and Z is a nucleotide sequence of 1 to 8 nucleotides in length,(b) is 100% complementary with a PRC2-associated region of a human SMN gene, wherein the PRC2-associated region is a region of the SMN gene that has a sequence that occurs at a higher frequency in a sequencing reaction of products of an RNA-immunoprecipitation assay that employs an antibody that targets Ezh2 to immunoprecipitate RNA-associated PRC2 complexes from cells comprising the SMN gene compared to a control sequencing reaction of products of a control RNA-immunoprecipitation assay that employs a control antibody, and(c) is 8 to 15 nucleotides in length, wherein, during the synthesis, at least one nucleotide incorporated into the oligonucleotide is a nucleotide analogue and/or a modified internucleotide linkage is incorporated between at least two nucleotides. 33. The method of claim 32, wherein delivery of the single stranded oligonucleotide into the cell results in a level of expression of SMN1 or SMN2 mRNA that is at least 50% greater than a level of expression of SMN1 or SMN2 mRNA in a control cell that does not comprise the single stranded oligonucleotide. 34. A method of increasing expression of SMN2 messenger RNA (mRNA) in a human cell, the method comprising: delivering to the cell a first single stranded oligonucleotide complementary with at least 8 consecutive nucleotides of a PRC2-associated region of human SMN2 and a second single stranded oligonucleotide complementary with at least 8 consecutive nucleotides of a splice control sequence of a precursor mRNA of human SMN2, in amounts sufficient to increase expression of a mature mRNA of SMN2 that comprises exon 7 in the cell, wherein the first and second oligonucleotides do not induce substantial cleavage or degradation of SMN2 mRNA in the cell. 35. The method of claim 34, wherein the first single stranded oligonucleotide is covalently linked to the second single stranded oligonucleotide through a linker. 36. A composition comprising: a first single stranded oligonucleotide produced by a process comprising synthesizing a single stranded oligonucleotide that is complementary with at least 8 consecutive nucleotides of a PRC2-associated region of a human SMN2 gene, wherein the PRC2-associated region is a region of the SMN2 gene that has a sequence that occurs at a higher frequency in a sequencing reaction of products of an RNA-immunoprecipitation assay that employs an antibody that targets Ezh2 to immunoprecipitate RNA-associated PRC2 complexes from cells comprising the SMN2 gene compared to a control sequencing reaction of products of a control RNA-immunoprecipitation assay that employs a control antibody, and a second single stranded oligonucleotide complementary with at least 9 consecutive nucleotides of a splice control sequence of a precursor mRNA encoded by the SMN2 gene. 37. The composition of claim 36, wherein the first single stranded oligonucleotide is covalently linked to the second single stranded oligonucleotide through a linker. 38. The composition of claim 36, wherein the first and/or single stranded oligonucleotide comprises at least one nucleotide analogue and/or a modified internucleotide linkage between at least two nucleotides.
연구과제 타임라인
LOADING...
LOADING...
LOADING...
LOADING...
LOADING...
이 특허에 인용된 특허 (74)
Cook Phillip Dan ; Guinosso Charles John, 2'-O-modified nucleosides and phosphoramidites.
Bennett C. Frank ; Cooke Stanley T. ; Manoharan Muthiah ; Wyatt Jacqueline R. ; Baker Brenda F. ; Monia Brett P. ; Freier Susan M. ; McKay Robert ; Karras James G., Alteration of cellular behavior by antisense modulation of mRNA processing.
David J. Ecker ; Phillip Dan Cook ; Brett P. Monia ; Susan M. Freier ; Yogesh S. Sanghvi, Antisense inhibition of ras gene with chimeric and alternating oligonucleotides.
Ecker David J. ; Cook Phillip Dan ; Monia Brett P. ; Freier Susan M. ; Sanghvi Yogesh S., Antisense inhibition of ras gene with chimeric and alternating oligonucleotides.
Cook, Phillip Dan; Manoharan, Muthiah; Bennett, Clarence Frank, Compositions and methods for enhanced biostability and altered biodistribution of oligonucleotides in mammals.
Walder Joseph A. ; Walder Roxanne Y. ; Eder Paul S. ; Dagle John M., DNA molecules stabilized by modifications of the 3'-terminal phosphodiester linkage.
Shay Jerry W. ; Wright Woodring E. ; Piatyszek Mieczyslaw A. ; Corey David R. ; Norton James C., Inhibition of mammalian telomerase by peptide nucleic acids.
Walder Joseph A. (Iowa City IA) Walder Roxanne Y. (Iowa City IA) Eder Paul S. (Iowa City IA) Dagle John M. (Iowa City IA), Methods for blocking the expression of specifically targeted genes.
Shay Jerry W. ; Wright Woodring E. ; Piatyszek Mieczyslaw A. ; Corey David R. ; Norton James C., Modulation of mammalian telomerase by peptide nucleic acids.
Shay Jerry W. ; Wright Woodring E. ; Piatyszek Mieczyslaw A. ; Corey David R. ; Norton James C., Modulation of mammalian telomerase by peptide nucleic acids.
Esau, Christine; Lollo, Bridget; Bennett, C. Frank; Freier, Susan M.; Griffey, Richard H.; Baker, Brenda F.; Vickers, Timothy A.; Marcusson, Eric G.; Koller, Erich; Swayze, Eric E.; Jain, Ravi; Bhat, Balkrishen; Peralta, Eigen, Oligomeric compounds and compositions for use in modulation of small non-coding RNAs.
Cook Phillip Dan (San Marcos CA) Hoke Glenn (Mt. Airy MD), Oligonucleotides for modulating Ha-ras or Ki-ras having phosphorothioate linkages of high chiral purity.
Thrue, Charlotte Albaek; Rosenbohm, Christoph; Hansen, Henrik Frydenlund; Westergaard, Majken; Mikkelsen, Nikolaj Dam; Christensen, Signe M.; Koch, Troels; Pedersen, Daniel Sejer; Frieden, Miriam, Oligonucleotides with alternating segments of locked and non-locked nucleotides.
Kauppinen, Sakari; Abrahamsen, Niels; Hildebrandt-Eriksen, Elisabeth S.; Munk, Martin, Pharmaceutical compositions for treatment of MicroRNA related diseases.
Ravikumar,Vasulinga T.; Manoharan,Muthiah; Capaldi,Daniel C.; Krotz,Achim; Cole,Douglas L.; Guzaev,Andrei, Process for the synthesis of oligomeric compounds.
Collard, Joseph; Khorkova Sherman, Olga; Coito, Carlos, Treatment of hemoglobin (HBF/HBG) related diseases by inhibition of natural antisense transcript to HBF/HBG.
※ AI-Helper는 부적절한 답변을 할 수 있습니다.