Selective oxidation of 5-methylcytosine by TET-family proteins
AssigneesChildren's Medical Center Corporation, The United States of America, As Reprsented by the Secretary, Department of Health & 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, TET3, and CXXC4. The novel activity is related to the enzymes being capable of converting the cytosine nucleotide 5-methylcytosine into 5-hydroxymethylcytosine by hydroxylation.
Claims26
A method for detecting a methylated cytosine residue in a mammalian nucleic acid, comprising:
- (a) contacting said mammalian nucleic acid comprising said methylated cytosine residue with a polypeptide having at least 90% sequence identity to:
- (i) a TET1 polypeptide;
- (ii) a TET2 polypeptide;
- (iii) a TET3 polypeptide;
- (iv) a CXXC4 polypeptide;
- (v) a catalytically active fragment of:
- (1) said TET1 polypeptide, (2) said TET2 polypeptide, (3) said TET3 polypeptide, or (4) said CXXC4 polypeptide;
- (vi) a variant thereof;
- or any combination thereof to generate a product nucleic acid comprising a modified methylated cytosine residue;
- (b) derivatizing said modified methylated cytosine residue in said product nucleic acid with a glucose molecule or a glucose-derivative donor substrate;
- and
- (c) performing a reaction on said product nucleic acid or a derivative thereof, to identify a presence of said methylated cytosine residue in said mammalian nucleic acid.
Dependencies are inferred from the claim text.