Selective oxidation of 5-methylcytosine by TET-family proteins
AssigneesChildren's Medical Center Corporation, The United States of America, as represented by the Secretary, Department of Health and Human Services
InventorsAnjana Rao, Mamta Tahiliani, Kian Peng Koh, Suneet Agarwal, Aravind Iyer
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Abstract
The present invention provides for novel methods for regulating and detecting the cytosine methylation status of DNA. The invention is based upon identification of a novel and surprising catalytic activity for the family of TET proteins, namely TET1, TET2, TEM3, and CXXC4. The novel activity is related to the enzymes being capable of converting the cytosine nucleotide 5-methylcytosine into 5-hydroxymethylcytosine by hydroxylation.
Claims30
A method comprising:
- (a) contacting a nucleic acid molecule comprising cytosine and 5-methylcytosine with an isolated enzyme comprising a TET polypeptide, a catalytically active fragment thereof, a variant thereof, or any combination thereof in an amount effective to convert 5-methylcytosine of said nucleic acid molecule to an oxidized derivative thereof;
- (b) converting cytosine of said nucleic acid molecule to uracil;
- and
- (c) conducting a sequencing reaction to obtain a sequence of said nucleic acid molecule.
A method comprising:
- contacting a nucleic acid molecule comprising cytosine and 5-methylcytosine with an isolated enzyme comprising a TET polypeptide, a catalytically active fragment thereof, a variant thereof, or any combination thereof in an amount effective to convert all of said 5-methylcytosine of said nucleic acid molecule to an oxidized derivative thereof.
Dependencies are inferred from the claim text.