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

被引：6
|
作者：
Tan, Darany [3 ]
Chung, Tzu Hung [3 ]
Sun, Xueguang [1 ,2 ]
Jia, Xi-Yu [3 ]
机构：
[1] Guangzhou Univ, Sch Life Sci, Guangzhou 510006, Guangdong, Peoples R China
[2] Cincinnati Childrens Hosp Med Ctr, Div Human Genet, 3333 Burnet Ave, Cincinnati, OH 45229 USA
[3] Zymo Res Corp, 17062 Murphy Ave, Irvine, CA 92614 USA
来源：
ANALYTICAL CHEMISTRY | 2018年 / 90卷 / 22期
关键词：
5-HYDROXYMETHYLCYTOSINE; DNA; 5-METHYLCYTOSINE;
D O I：
10.1021/acs.analchem.8b02832
中图分类号：
O65 [分析化学];
学科分类号：
070302 ; 081704 ;
摘要：
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 beta-glucosyltrans-ferase 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.
引用
收藏
页码：13200 / 13206
页数：7
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