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- Title
Selective chemical labeling reveals the genome-wide distribution of 5-hydroxymethylcytosine.
- Authors
Chun-Xiao Song; Szulwach, Keith E.; Ye Fu; Qing Dai; Chengqi Yi; Xuekun Li; Yujing Li; Chih-Hsin Chen; Wen Zhang; Xing Jian; Jing Wang; Li Zhang; Looney, Timothy J.; Baichen Zhang; Godley, Lucy A.; Hicks, Leslie M.; Lahn, Bruce T.; Peng Jin; Chuan He
- 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.
- Publication
Nature Biotechnology, 2011, Vol 29, Issue 1, p68
- ISSN
1087-0156
- Publication type
Academic Journal
- DOI
10.1038/nbt.1732