CCCTC-binding factor (CTCF) RNA interactome
This invention relates to methods and compositions for selectively reactivating or downregulating certain genes, e.g., genes regulated by zinc-finger protein CCCTC-binding factor (CTCF) on autosomes (e.g., imprinted genes, tumor suppressors, cancer) and the inactive X chromosome (Xi), e.g., genes associated with X-linked diseases, e.g., Rett Syndrome, Factor VIII or IX deficiency, Fragile X Syndrome, Duchenne muscular dystrophy, and PNH, in heterozygous females carrying a mutated allele, in addition to a functional wildtype or hypomorphic allele.
1. A method of activating an inactive X-linked allele in a cell, preferably a cell of a female heterozygous subject, the method comprising administering to the cell an inhibitory oligonucleotide targeting a sequence of at least 12 consecutive nucleotides within SEQ ID NOs:49590-49613, wherein CTCF binding to Jpx RNA is disrupted.
2. The method of claim 1 , wherein the inactive X-linked allele is associated with an X-linked disorder, and the oligonucleotide is administered in a therapeutically effective amount.
3. The method of claim 1 , wherein the cell is in a living subject.
4. The method of claim 1 , wherein the inhibitory oligonucleotide is identical or complementary to at least 15 consecutive nucleotides of SEQ ID NOs:49590-49613.
5. The method of claim 1 , wherein the oligonucleotide does not comprise three or more consecutive guanosine nucleotides.
6. The method of claim 1 , wherein the oligonucleotide does not comprise four or more consecutive guanosine nucleotides.
7. The method of claim 1 , wherein the oligonucleotide is 12 to 30 nucleotides in length.
8. The method of claim 1 , wherein at least one nucleotide of the oligonucleotide is a nucleotide analogue or a 2′ O-methyl.
9. The method of claim 1 , wherein the oligonucleotide comprises at least one ribonucleotide, at least one deoxyribonucleotide, or at least one bridged nucleotide.
10. The method of claim 9 , wherein the bridged nucleotide is a LNA nucleotide, a cEt nucleotide or a ENA modified nucleotide.
11. The method of claim 1 , wherein one or more of the nucleotides of the oligonucleotide comprise 2′-fluoro-deoxyribonucleotides, one or more of the nucleotides of the oligonucleotide comprise 2′-O-methyl nucleotides, one or more of the nucleotides of the oligonucleotide comprise ENA nucleotide analogues, and/or one or more of the nucleotides of the oligonucleotide comprise LNA nucleotides.
12. The method of claim 1 , wherein the nucleotides of the oligonucleotide comprise comprising phosphorothioate internucleotide linkages between at least two nucleotides or between all nucleotides.
13. The method of claim 1 , wherein the nucleotides of the oligonucleotide comprise phosphorothioate internucleotide linkages between at least two nucleotides.
14. The method of claim 1 , wherein the nucleotides of the oligonucleotide comprise phosphorothioate internucleotide linkages between all nucleotides.
15. The method of claim 1 , wherein the oligonucleotide is a gapmer or a mixmer.
16. The method of claim 1 , wherein the inactive X-linked allele is CDKL5, MECP2, or FMR1.