Composition and methods related to modification of 5-hydroxymethylcytosine (5-hmC)
First Claim
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1. 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.
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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.
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Citations
25 Claims
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1. 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.- View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 19)
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17. A method for mapping by glycosylation with a modified glucose molecule 5-hydroxymethylcytosine in a nucleic acid molecule of an isolated sample comprising:
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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; andmapping the glycosylated 5-hydroxymethylcytosines in the glycosylated nucleic acid molecule by sequencing the glycosylated nucleic acid molecule. - View Dependent Claims (18, 20, 21, 22, 23, 24, 25)
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Specification