Mirror Bisulfite Sequencing: A Method for Single-Base Resolution of Hydroxymethylcytosine | Zendy

Darany Tan | Zendy; Tzu Hung Chung | Zendy; Xueguang Sun | Zendy; Xi-Yu Jia | Zendy

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Mirror Bisulfite Sequencing: A Method for Single-Base Resolution of Hydroxymethylcytosine

Author(s) -

Darany Tan,

Tzu Hung Chung,

Xueguang Sun,

Xi-Yu Jia

Publication year - 2018

Publication title -

analytical chemistry

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 2.117

H-Index - 332

eISSN - 1520-6882

pISSN - 0003-2700

DOI - 10.1021/acs.analchem.8b02832

Subject(s) - 5 hydroxymethylcytosine , chemistry , 5 methylcytosine , cpg site , semiconservative replication , bisulfite sequencing , cytosine , dna , bisulfite , base pair , dna methylation , microbiology and biotechnology , biochemistry , biology , gene , plasmid , origin of replication , gene expression

Although the role of 5-methylcytosine has been well studied, the biological role of 5-hydroxymethylcytosine still remains unclear because of the limited methods available for single-base detection of 5-hydroxymethylcytosine (5hmC). Here, we present mirror bisulfite sequencing for 5hmC detection at a single CpG site by synthesizing a DNA strand to mirror the parental strand. This semiconservative duplex is sequentially treated with β-glucosyltransferase and M.SssI methylase. The glucosyl-5hmCpG in the parental strand inhibits methylation of the mirroring CpG site, and after bisulfite conversion, a thymine in the mirroring strand indicates a 5hmCpG site in the parental strand, whereas a cytosine indicates a non-5hmC site. Using this method, the 5hmC levels of various human tissues and paired liver tissues were mapped genomewide.

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