Hsu Hang-Kai, Weng Yu-I, Hsu Pei-Yin, Huang Tim H-M, Huang Yi-Wen
The Ohio State University Comprehensive Cancer Center, Columbus, OH, USA.
Methods Mol Biol. 2014;1105:61-70. doi: 10.1007/978-1-62703-739-6_5.
DNA methylation has been characterized as the representative example of epigenetic modifications and implicated in numerous biological processes, such as genomic imprinting and X chromosome inactivation. It primarily occurs at CpG dinucleotides in mammals and plays a critical role in transcriptional regulations. Examination of DNA methylation patterns in gene(s) or across a genome is vital to understand the role of epigenetic modulation in the progress of development and tumorigenesis. Currently, lots of approaches have been developed to investigate DNA methylation patterns for either limited regions or for genome-scale studies, but some of them rely on using restriction enzymes. In this chapter, we describe two commonly used protocols to detect enrichment of methylated DNA regions, namely, methylated DNA immunoprecipitation (MeDIP) and capture of methylated DNA by methyl-CpG binding domain-based (MBD) proteins (MBDCap). They are the most economical and effective methods to study DNA methylation either at a single locus or in genome-wide scale.
DNA甲基化已被视为表观遗传修饰的典型例子，并与众多生物学过程相关，如基因组印记和X染色体失活。它主要发生在哺乳动物的CpG二核苷酸处，在转录调控中起关键作用。检测基因或全基因组中的DNA甲基化模式对于理解表观遗传调控在发育和肿瘤发生过程中的作用至关重要。目前，已经开发了许多方法来研究有限区域或全基因组规模的DNA甲基化模式，但其中一些方法依赖于使用限制性酶。在本章中，我们描述了两种常用的检测甲基化DNA区域富集的方案，即甲基化DNA免疫沉淀（MeDIP）和基于甲基-CpG结合域（MBD）蛋白捕获甲基化DNA（MBDCap）。它们是在单个位点或全基因组范围内研究DNA甲基化最经济有效的方法。
