METHODS OF EDITING DNA METHYLATION
The invention relates to methods of modifying DNA methylation by contacting a cell with a catalytically inactive site specific nuclease fused to an effector domain having methylation or demethylation activity and one or more guide sequences.
1 . A method of treating a patient in need thereof, the method comprising administering to the patient:
a) a nucleic acid that encodes a polypeptide comprising a catalytically inactive site specific nuclease fused to an effector domain have methylation activity; and
b) a guide sequence or a nucleic acid that encodes a guide sequence.
2 . The method of claim 1 , wherein the guide sequence targets the polypeptide to one or more genomic sequences.
3 . The method of claim 2 , wherein one or more genomic sequences are aberrantly hypomethylated or aberrantly hypermethylated.
4 . The method of claim 2 , wherein the subject has a disease or disorder associated with or produced by a genetic modification in one or more genomic sequences.
5 . The method of claim 4 , wherein the one or more genomic sequences are involved in pathogenesis of the disease or disorder.
6 . The method of claim 2 , wherein the one or more genomic sequences comprise a CTCF binding site.
7 . The method of claim 1 , wherein the patient has a cancer.
8 . The method of claim 1 , wherein the patient has an inflammatory disease or disorder.
9 . A method of treating a disease or disorder in a subject comprising administering to the subject:
a) a polypeptide comprising a catalytically inactive site specific nuclease fused to an effector domain having methylation or demethylation activity, or a nucleic acid encoding the polypeptide; and
b) a guide sequence or a nucleic acid that encodes a guide sequence, wherein the guide sequence targets a genomic sequence comprising a CTCF binding site.
10 . The method of claim 9 , wherein the genomic sequence is aberrantly hypomethylated or aberrantly hypermethylated.
11 . The method of claim 9 , wherein the disease or disorder is associated with or produced by a genetic modification in the genomic sequence.
12 . The method of claim 9 , wherein the genomic sequence is involved in pathogenesis of the disease or disorder.
13 . The method of claim 9 , wherein the disease or disorder is a cancer.
14 . The method of claim 9 , wherein the disease or disorder is an inflammatory disease or disorder.
15 . The method of claim 9 , wherein the effector domain comprises Tet1 or Dnmt3a.
16 . The method of claim 9 , wherein the catalytically inactive site specific nuclease is a catalytically inactive Cas protein.
17 . The method of claim 9 , wherein the catalytically inactive site specific nuclease is a catalytically inactive Cas9 protein.
18 . The method of claim 9 , wherein the catalytically inactive site specific nuclease is a catalytically inactive Cpf1 protein.
19 . The method of claim 9 , wherein the guide sequence is a ribonucleic acid guide sequence.
20 . The method of claim 9 , wherein the guide sequence is from about 10 base pairs to about 150 base pairs in length.