Quantitation and mapping of the epigenetic marker 5-hydroxymethylcytosine.
We here review primary methods used in quantifying and mapping 5-hydroxymethylcytosine (5hmC), including global quantification, restriction enzyme-based detection, and methods involving DNA-enrichment strategies and the genome-wide sequencing of 5hmC. As discovered in the mammalian genome in 2009, 5hmC, oxidized from 5-methylcytosine (5mC) by ten-eleven translocation (TET) dioxygenases, is increasingly being recognized as a biomarker in biological processes from development to pathogenesis, as its various detection methods have shown. We focus in particular on an ultrasensitive single-molecule imaging technique that can detect and quantify 5hmC from trace samples and thus offer information regarding the distance-based relationship between 5hmC and 5mC when used in combination with fluorescence resonance energy transfer.
Department of Cancer Biology, University of Cincinnati College of Medicine, Cincinnati, OH, USA.
Ying
Y
Qing
Department of Cancer Biology, University of Cincinnati College of Medicine, Cincinnati, OH, USA.|Center for Mitochondrial Biology and Medicine, The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology Xi'an Jiaotong University, Xi'an, China.
Zhiqi
Z
Tian
Ludwig Institute for Cancer Research, Nuffield Department of Medicine, University of Oxford, Oxford, UK.
Ying
Y
Bi
Department of Cancer Biology, University of Cincinnati College of Medicine, Cincinnati, OH, USA.|Center for Mitochondrial Biology and Medicine, The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology Xi'an Jiaotong University, Xi'an, China.
Yongyao
Y
Wang
Center for Mitochondrial Biology and Medicine, The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology Xi'an Jiaotong University, Xi'an, China.
Jiangang
J
Long
Ludwig Institute for Cancer Research, Nuffield Department of Medicine, University of Oxford, Oxford, UK.|Target Discovery Institute, Nuffield Department of Medicine, University of Oxford, Oxford, UK.
Chun-Xiao
CX
Song
Department of Cancer Biology, University of Cincinnati College of Medicine, Cincinnati, OH, USA.
Jiajie
0000-0003-4288-3203
J
Diao
D044503:5-Methylcytosine / Q000031:analogs & derivatives* / Q000032:analysis / Q000276:immunology / Q000378:metabolism; D000818:Animals; D000918:Antibody Specificity; D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D002874:Chromosome Mapping; D004247:DNA / Q000737:chemistry / Q000235:genetics; D004262:DNA Restriction Enzymes; D044127:Epigenesis, Genetic; D031541:Fluorescence Resonance Energy Transfer; D005819:Genetic Markers; D006031:Glycosylation; D006801:Humans; D013058:Mass Spectrometry; D017422:Sequence Analysis, DNA; D000072760:Single Molecule Imaging / Q000379:methods
D005819:Genetic Markers; C011865:5-hydroxymethylcytosine; D044503:5-Methylcytosine; D004247:DNA; D004262:DNA Restriction Enzymes
 single molecule imaging; epigenetic modification