MBD-isolated Genome Sequencing provides a high-throughput and comprehensive survey of DNA methylation in the human genome

被引:280
作者
Serre, David [1 ]
Lee, Byron H. [2 ]
Ting, Angela H. [1 ]
机构
[1] Cleveland Clin Fdn, Lerner Res Inst, Genom Med Inst, Cleveland, OH 44195 USA
[2] Cleveland Clin Fdn, Glickman Urol & Kidney Inst, Cleveland, OH 44195 USA
关键词
HUMAN CANCER-CELLS; ABERRANT METHYLATION; GENE; PROMOTER; HYPERMETHYLATION; INACTIVATION; EPITHELIA; METHYLOME; BREAST;
D O I
10.1093/nar/gkp992
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
DNA methylation is an epigenetic modification involved in both normal developmental processes and disease states through the modulation of gene expression and the maintenance of genomic organization. Conventional methods of DNA methylation analysis, such as bisulfite sequencing, methylation sensitive restriction enzyme digestion and array-based detection techniques, have major limitations that impede high-throughput genome-wide analysis. We describe a novel technique, MBD-isolated Genome Sequencing (MiGS), which combines precipitation of methylated DNA by recombinant methyl-CpG binding domain of MBD2 protein and sequencing of the isolated DNA by a massively parallel sequencer. We utilized MiGS to study three isogenic cancer cell lines with varying degrees of DNA methylation. We successfully detected previously known methylated regions in these cells and identified hundreds of novel methylated regions. This technique is highly specific and sensitive and can be applied to any biological settings to identify differentially methylated regions at the genomic scale.
引用
收藏
页码:391 / 399
页数:9
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