Polycomb-associated non-coding RNAs

Patent metadata

Field	Value
Publication number	US-11066673-B2
Application number	US-201615265104-A
Country	US
Kind code	B2
Filing date	Sep 14, 2016
Priority date	Nov 12, 2010
Publication date	Jul 20, 2021
Grant date	Jul 20, 2021

Abstract

This invention relates to long non-coding RNAs (lncRNAs), libraries of those lncRNAs 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, and wherein recruitment of PRC2 to the target gene by the PRC2-binding RNA results in an increase in trimethylation of histone H3-lysine27 at the target gene, the method comprising delivering to the cell a composition comprising an isolated oligonucleotide mimetic that specifically interferes with binding of PRC2 to a PRC2-binding region of the PRC2-binding RNA without inducing degradation of the PRC2-binding RNA by RNAse H, 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, and wherein a decrease in recruitment of PRC2 to the target gene in the cell and a decrease in trimethylation of histone H3-lysine27 at the target gene following delivery of the oligonucleotide mimetic to the cell, compared with an appropriate control cell to which the oligonucleotide mimetic has not been delivered, indicates effectiveness of the oligonucleotide mimetic.

2. The method of claim 1 , further comprising detecting expression of the PRC2-binding RNA in the cell, wherein expression of the PRC2-binding RNA in the cell indicates that the oligonucleotide mimetic is suitable for increasing expression of the target gene in the cell.

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

4. The method of claim 1 , wherein the oligonucleotide mimetic is a peptide nucleic acid (PNA) compound.

5. The method of claim 4 , wherein the PNA compound comprises an aminoethylglycine backbone.

6. The method of claim 1 , wherein the oligonucleotide mimetic is a locked nucleic acid (LNA) compound.

7. The method of claim 1 , wherein the oligonucleotide mimetic is a morpholino-based oligomeric compound.

8. The method of claim 7 , wherein the morpholino-based oligomeric compound is a phosphorodiamidate morpholino oligomer (PMO).

9. The method of claim 1 , wherein the oligonucleotide mimetic is a cyclohexenyl nucleic acid compound.

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

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

12. The method of claim 1 , wherein the cell is a human cell.

13. The method of claim 1 , wherein the cell is a cancer cell.

14. The method of claim 1 , wherein the cell is a muscle cell.

15. The method of claim 1 , wherein the cell is a liver cell.

16. The method of claim 11 , wherein the PRC2-binding RNA is transcribed from the same strand of the chromosomal region as the target gene.

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

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

19. The method of claim 1 , wherein the antibody directed against PRC2 is an antibody that selectively binds to EZH2.

20. The method of claim 1 , wherein the antibody directed against PRC2 is an antibody that selectively binds to Eed, Suz12 or RbAp48.

21. 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, and wherein recruitment of PRC2 to the target gene by the PRC2-binding RNA results in an increase in trimethylation of histone H3-lysine27 at the target gene, the method comprising delivering to the cell a composition comprising a means for specifically inhibiting binding of PRC2 to a PRC2-binding region of the PRC2-binding RNA to increase expression of the target gene without inducing degradation of the PRC2-binding RNA by RNAse H, 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, and wherein a decrease in recruitment of PRC2 to the target gene in the cell and a decrease in trimethylation of histone H3-lysine27 at the target gene following delivery of the composition to the cell, compared with an appropriate control cell to which the composition has not been delivered, indicates effectiveness of the composition.

Assignees

• Massachusetts Gen Hospital

Inventors

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

Classifications

• C12N15/113

  Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; {Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing (when used in plants C12N15/8218)} · CPC title

• C12N2310/346

  having a combination of backbone and sugar modifications · CPC title

• C12N2310/314

  Phosphoramidates · CPC title

• C12N2310/321

  2'-O-R Modification · CPC title

• A61P13/00

  Drugs for disorders of the urinary system · CPC title