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, 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 of converting a methylated cytosine residue in an isolated eukaryotic nucleic acid molecule to a modified cytosine residue, said method comprising:
- (a) contacting said isolated eukaryotic nucleic acid molecule with a polypeptide that converts said methylated cytosine residue to said modified cytosine residue,
- wherein said polypeptide comprises a TET1 polypeptide, a TET2 polypeptide, a TET3 polypeptide, or a CXXC4 polypeptide, or a catalytically active fragment thereof, and
- (b) contacting said isolated eukaryotic nucleic acid molecule with a glucosyltransferase polypeptide to derivatize said modified cytosine residue, wherein said glucosyltransferase polypeptide comprises β-glucosyltransferase (BGT) or α-glucosyltransferase (AGT).
Dependencies are inferred from the claim text.