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Nat Biotechnol. 2011 Jan;29(1):68-72. doi: 10.1038/nbt.1732. Epub 2010 Dec 12.

Selective chemical labeling reveals the genome-wide distribution of 5-hydroxymethylcytosine.

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  • 1Department of Chemistry and Institute for Biophysical Dynamics, the University of Chicago, Chicago, Illinois, USA.

Abstract

In contrast to 5-methylcytosine (5-mC), which has been studied extensively, little is known about 5-hydroxymethylcytosine (5-hmC), a recently identified epigenetic modification present in substantial amounts in certain mammalian cell types. Here we present a method for determining the genome-wide distribution of 5-hmC. We use the T4 bacteriophage β-glucosyltransferase to transfer an engineered glucose moiety containing an azide group onto the hydroxyl group of 5-hmC. The azide group can be chemically modified with biotin for detection, affinity enrichment and sequencing of 5-hmC-containing DNA fragments in mammalian genomes. Using this method, we demonstrate that 5-hmC is present in human cell lines beyond those previously recognized. We also find a gene expression level-dependent enrichment of intragenic 5-hmC in mouse cerebellum and an age-dependent acquisition of this modification in specific gene bodies linked to neurodegenerative disorders.

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PMID:
21151123
[PubMed - indexed for MEDLINE]
PMCID:
PMC3107705
Free PMC Article

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