SELECTIVE OXIDATION OF 5-METHYLCYTOSINE BY TET-FAMILY PROTEINS
First Claim
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1. A method comprising:
- contacting a mammalian nucleic acid sequence with an enzyme that utilizes a labeled glucose or a labeled glucose-derivative donor substrate to add a labeled glucose molecule or a labeled glucose-derivative to a 5-hydroxymethylcytosine in said mammalian nucleic acid sequence to generate a labeled glucosylated-5-hydroxymethylcytosine, wherein said enzyme comprises a glucosyltransferase and wherein said labeled glucose or said labeled glucose-derivative donor substrate comprises a uridine diphosphate glucose (UDPG).
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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.
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14 Claims
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1. A method comprising:
contacting a mammalian nucleic acid sequence with an enzyme that utilizes a labeled glucose or a labeled glucose-derivative donor substrate to add a labeled glucose molecule or a labeled glucose-derivative to a 5-hydroxymethylcytosine in said mammalian nucleic acid sequence to generate a labeled glucosylated-5-hydroxymethylcytosine, wherein said enzyme comprises a glucosyltransferase and wherein said labeled glucose or said labeled glucose-derivative donor substrate comprises a uridine diphosphate glucose (UDPG). - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14)
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