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US 8,741,567 B2Granted
Composition and methods related to modification of 5-hydroxymethylcytosine (5-hmC)
AssigneeThe University of Chicago
InventorsChuan He, Chunxiao Song
Life of this patent
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Abstract
The present invention relates generally to the field of molecular biology. More particularly, it concerns methods and compositions for detecting, evaluating, and/or mapping 5-hydroxymethyl-modified cytosine bases within a nucleic acid molecule.
Claims25
A method for distinguishing by glycosylation with a modified glucose molecule 5-hydroxymethylcytosine from 5-methylcytosine in a nucleic acid molecule of an isolated sample comprising:
- placing in a reaction mixture the nucleic acid molecule with β-glucosyltransferase and a synthetically modified uridine diphosphoglucose (UDP-Glu) molecule comprising a modification moiety to glycosylate 5-hydroxymethyl-cytosines in the nucleic acid molecule with a modified glucose molecule, wherein the modification moiety comprises a linker, a detectable moiety, an isolation tag, a blocking moiety, or a functional moiety.
A method for mapping by glycosylation with a modified glucose molecule 5-hydroxymethylcytosine in a nucleic acid molecule of an isolated sample comprising:
- placing in a reaction mixture the nucleic acid molecule with β-glucosyltransferase and a synthetically modified uridine diphosphoglucose (UDP-Glu) molecule comprising a modification moiety to glycosylate 5-hydroxymethylcytosines in the nucleic acid molecule with the modified Glu molecule, wherein the modification moiety comprises a linker, a detectable moiety, an isolation tag, a blocking moiety, or a functional moiety;
- and
- mapping the glycosylated 5-hydroxymethylcytosines in the glycosylated nucleic acid molecule by sequencing the glycosylated nucleic acid molecule.
Dependencies are inferred from the claim text.