造血细胞中DNA甲基化的全基因组分析：采用全基因组亚硫酸氢盐测序法进行DNA甲基化分析

Genome-Wide Analysis of DNA Methylation in Hematopoietic Cells: DNA Methylation Analysis by WGBS.

作者信息

Jeong Mira, Guzman Anna G, Goodell Margaret A

机构信息

Department of Pediatrics, Baylor College of Medicine and Texas Children's Hospital, Houston, TX, USA.

Dan L. Duncan Cancer Center, Department of Molecular and Human Genetics, Stem Cells and Regenerative Medicine Center, and Center for Cell and Gene Therapy, Baylor College of Medicine, Houston, TX, USA.

出版信息

Methods Mol Biol. 2017;1633:137-149. doi: 10.1007/978-1-4939-7142-8_9.

DOI:10.1007/978-1-4939-7142-8_9

PMID:28735485

Abstract

DNA methylation is a major epigenetic modification that regulates gene expression, genome imprinting, and development and has a role in diseases including cancer. There are various methods for whole-genome methylation profiling that differ in cost and resolution. Recent advances in high-throughput sequencing technologies coupled with bisulfite treatment enable absolute DNA methylation quantification and genome-wide single-nucleotide resolution analysis. In this chapter, we provide detailed protocols for whole-genome bisulfite sequencing (WGBS), which captures the complete methylome. Using WGBS, we are able to generate a reference DNA methylome for normal or malignant hematopoietic cells.

摘要

DNA甲基化是一种主要的表观遗传修饰，可调节基因表达、基因组印记和发育，并在包括癌症在内的疾病中发挥作用。全基因组甲基化谱分析有多种方法，这些方法在成本和分辨率上有所不同。高通量测序技术与亚硫酸氢盐处理的最新进展使得能够进行绝对DNA甲基化定量和全基因组单核苷酸分辨率分析。在本章中，我们提供了全基因组亚硫酸氢盐测序（WGBS）的详细方案，该方法可捕获完整的甲基化组。使用WGBS，我们能够生成正常或恶性造血细胞的参考DNA甲基化组。