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Single base resolution analysis of 5-hydroxymethylcytosine in 188 human genes: implications for hepatic gene expression
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Number of Authors: 10
2016 (English)In: Nucleic Acids Research, ISSN 0305-1048, E-ISSN 1362-4962, Vol. 44, no 14, 6756-6769 p.Article in journal (Refereed) Published
Abstract [en]

To improve the epigenomic analysis of tissues rich in 5-hydroxymethylcytosine (hmC), we developed a novel protocol called TAB-Methyl-SEQ, which allows for single base resolution profiling of both hmC and 5-methylcytosine by targeted next-generation sequencing. TAB-Methyl-SEQ data were extensively validated by a set of five methodologically different protocols. Importantly, these extensive cross-comparisons revealed that protocols based on Tet1-assisted bisulfite conversion provided more precise hmC values than TrueMethyl-based methods. A total of 109 454 CpG sites were analyzed by TAB-Methyl-SEQ for mC and hmC in 188 genes from 20 different adult human livers. We describe three types of variability of hepatic hmC profiles: (i) sample-specific variability at 40.8% of CpG sites analyzed, where the local hmC values correlate to the global hmC content of livers (measured by LC-MS), (ii) gene-specific variability, where hmC levels in the coding regions positively correlate to expression of the respective gene and (iii) site-specific variability, where prominent hmC peaks span only 1 to 3 neighboring CpG sites. Our data suggest that both the gene-and site-specific components of hmC variability might contribute to the epigenetic control of hepatic genes. The protocol described here should be useful for targeted DNA analysis in a variety of applications.

Place, publisher, year, edition, pages
2016. Vol. 44, no 14, 6756-6769 p.
National Category
Biological Sciences
URN: urn:nbn:se:su:diva-134419DOI: 10.1093/nar/gkw316ISI: 000382999900024PubMedID: 27131363OAI: diva2:1037244
Available from: 2016-10-14 Created: 2016-10-06 Last updated: 2016-10-14Bibliographically approved

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Ewels, Philip
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Department of Biochemistry and BiophysicsScience for Life Laboratory (SciLifeLab)
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