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 Servic
InventorsAnjana Rao, Mamta Tahiliani, Kian Peng Koh, Suneet Agarwal, Aravind Iyer
Life of this patent
Estimating the term…
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.
Claims27
A method comprising:
- (a) contacting a mammalian nucleic acid sequence with an endonuclease, wherein said endonuclease cleaves nucleic acid sequences at a recognition site comprising a hydroxymethylated base;
- and
- (b) detecting cleavage of said mammalian nucleic acid sequence to identify said hydroxymethylated base.
A method comprising:
- (a) contacting a mammalian nucleic acid sequence with an endonuclease, wherein said mammalian nucleic acid sequence comprises a hydroxymethylated base;
- and
- (b) distinguishing methylated base and hydroxymethylated base in said mammalian nucleic acid sequence based on cleavage activity of said endonuclease.
A method comprising:
- (a) modifying a hydroxymethylated base in a nucleic acid sequence using a glucosyltransferase;
- and
- (b) contacting said nucleic acid sequence with an endonuclease, wherein said endonuclease cleaves nucleic acid sequences at a recognition site comprising a hydroxymethylated base or a methylated base.
Dependencies are inferred from the claim text.