Polycomb-associated non-coding RNAs

Patent metadata

Field	Value
Publication number	US-9816094-B2
Application number	US-201615171706-A
Country	US
Kind code	B2
Filing date	Jun 2, 2016
Priority date	Nov 12, 2010
Publication date	Nov 14, 2017
Grant date	Nov 14, 2017

Abstract

This invention relates to long non-coding RNAs (lncRNAs), libraries of those ncRNAs that bind chromatin modifiers, such as Polycomb Repressive Complex 2, inhibitory nucleic acids and methods and compositions for targeting lncRNAs.

First claim

What is claimed is:

1. A method of inducing expression of a non-imprinted target gene in a cell, wherein expression of the target gene is inhibited by a PRC2-binding RNA expressed in target cells, the method comprising:
delivering to the cell a composition comprising an isolated single stranded oligonucleotide of 8 to 25 nucleotides in length having a region of complementarity that is complementary with at least 5 contiguous nucleotides of the PRC2-binding RNA that inhibits expression of the target gene, wherein the oligonucleotide is complementary to and binds specifically within a PRC2-binding region of the PRC2-binding RNA and interferes with binding of PRC2to the PRC2-binding region without inducing degradation of the PRC2-binding RNA, wherein the PRC2-binding region has a nucleotide sequence protected from nucleases during an RNA immunoprecipitation procedure using an antibody directed against PRC2, wherein the PRC2-binding RNA is not ANRIL lncRNA and is transcribed from a sequence of the chromosomal locus of the target gene, wherein the single stranded oligonucleotide comprises at least one nucleotide having a 2′-O-methoxyethyl modified sugar moiety and/or at least one ribonucleic acid analogue having a ribose ring having a bridge between its 2′-oxygen and 4′-carbon, and wherein a decrease in recruitment of PRC2to the target gene in the cell following delivery of the single stranded oligonucleotide to the cell, compared with an appropriate control cell to which the single stranded oligonucleotide has not been delivered, indicates effectiveness of the single stranded oligonucleotide.

2. The method of claim 1 , wherein at least one nucleotide of the oligonucleotide has a 2′-O-methoxyethyl modified sugar moiety.

3. The method of claim 1 , wherein at least one nucleotide of the oligonucleotide is a ribonucleic acid analogue comprising a ribose ring having a bridge between its 2′-oxygen and 4′-carbon.

4. The method of claim 3 , wherein the ribonucleic acid analogue comprises a methylene bridge between the 2′-oxygen and the 4′-carbon.

5. The method of claim 1 , wherein at least one nucleotide of the oligonucleotide has a 2′-O-methoxyethyl modified sugar moiety and at least one nucleotide of the oligonucleotide is a ribonucleic acid analogue comprising a ribose ring having a bridge between its 2′-oxygen and 4′-carbon.

6. The method of claim 1 , wherein the oligonucleotide comprises at least one modified internucleoside linkage.

7. The method of claim 6 , wherein the at least one modified internucleoside linkage is selected from phosphorothioate, phosphorodithioate, alkylphosphonothioate, phosphoramidate, carbamate, carbonate, phosphate triester, acetamidate, carboxymethyl ester, and combinations thereof.

8. The method of claim 2 , wherein the oligonucleotide comprises at least one modified internucleoside linkage.

9. The method of claim 8 , wherein the at least one modified internucleoside linkage is a phosphorothioate.

10. The method of claim 3 , wherein the oligonucleotide comprises at least one modified internucleoside linkage.

11. The method of claim 10 , wherein the at least one modified internucleoside linkage is a phosphorothioate.

12. The method of claim 5 , wherein the oligonucleotide comprises at least one modified internucleoside linkage.

13. The method of claim 12 , wherein the at least one modified internucleoside linkage is a phosphorothioate.

14. The method of claim 1 , wherein at least one nucleotide of the oligonucleotide has a 2′-O-methoxyethyl modified sugar moiety and at least one nucleotide of the oligonucleotide is a ribonucleic acid analogue comprising a ribose ring having a bridge between its 2′-oxygen and 4′-carbon, and wherein each internucleoside linkage of the oligonucleotide is a modified internucleoside linkage.

15. The method of claim 14 , wherein the modified internucleoside linkage is a phosphorothioate.

16. The method of claim 1 , wherein at least one nucleotide of the oligonucleotide is an RNA or DNA nucleotide.

17. The method of claim 1 , wherein the RNA is a lncRNA.

18. The method of claim 1 , further comprising detecting expression of the PRC2-binding RNA in the cell before delivering to the cell a composition comprising the isolated single stranded oligonucleotide, wherein expression of the PRC2-binding RNA in the cell indicates that the single stranded oligonucleotide is suitable for increasing expression of the target gene in the cell.

19. The method of claim 1 , further comprising detecting a change in expression of the target gene following delivery of the single stranded oligonucleotide to the cell, wherein an increase in expression of the target gene compared with an appropriate control cell indicates effectiveness of the single stranded oligonucleotide.

20. The method of claim 1 , wherein the cell is in vitro.

21. The method of claim 1 , wherein the cell is in vivo.

22. The method of claim 1 , wherein the PRC2-binding RNA is transcribed from the same strand of the chromosomal region as the target gene.

23. The method of claim 1 , wherein the target gene is a protein-coding gene.

24. The method of claim 1 , wherein the chromosomal locus of the target gene is a gene of an autosomal chromosome.

25. The method of claim 1 , wherein the oligonucleotide has complementarity to a region of the PRC2-binding RNA transcribed from a portion of the target gene corresponding to an intron-exon junction or an intron.

26. The method of claim 1 , wherein the oligonucleotide has complementarity to a region of the PRC2-binding RNA transcribed from a portion of the target gene corresponding to an exon.

27. The method of claim 1 , wherein the oligonucleotide has complementarity to a region of the PRC2-binding RNA transcribed from a portion of the target gene corresponding to a translation initiation region or a translation termination region.

28. The method of claim 1 , wherein the oligonucleotide has complementarity to a region of the PRC2-binding RNA transcribed from a portion of the target gene corresponding to a promoter.

29. The method of claim 1 , wherein the oligonucleotide has complementarity to a region of the PRC2-binding RNA transcribed from a portion of the target gene corresponding to a 5′-UTR.

30. The method of claim 1 , wherein the oligonucleotide has complementarity to a region of the PRC2-binding RNA transcribed from a portion of the target gene corresponding to a 3′-UTR.

Assignees

• Massachusetts Gen Hospital

Inventors

• Lee Jeannie T
• Zhao Jing
• Sarma Kavitha
• Borowsky Mark
• Ohsumi Toshiro Kendrick

Classifications

• A61P9/00

  Drugs for disorders of the cardiovascular system · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• A61P3/00

  Drugs for disorders of the metabolism (of the blood or the extracellular fluid A61P7/00) · CPC title

• A61P29/00

  Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID] · CPC title

• A61P13/00

  Drugs for disorders of the urinary system · CPC title