Department of Biology, University of Alabama at Birmingham, Birmingham, AL, United States.
Department of Biology, University of Alabama at Birmingham, Birmingham, AL, United States; Comprehensive Cancer Center, University of Alabama at Birmingham, Birmingham, AL, United States; Nutrition Obesity Research Center, University of Alabama at Birmingham, Birmingham, AL, United States; Comprehensive Center for Healthy Aging, University of Alabama at Birmingham, Birmingham, AL, United States; Comprehensive Diabetes Center, University of Alabama at Birmingham, Birmingham, AL, United States.
Methods. 2021 Mar;187:28-43. doi: 10.1016/j.ymeth.2020.10.002. Epub 2020 Oct 9.
DNA methylation provides a pivotal layer of epigenetic regulation in eukaryotes that has significant involvement for numerous biological processes in health and disease. The function of methylation of cytosine bases in DNA was originally proposed as a "silencing" epigenetic marker and focused on promoter regions of genes for decades. Improved technologies and accumulating studies have been extending our understanding of the roles of DNA methylation to various genomic contexts including gene bodies, repeat sequences and transcriptional start sites. The demand for comprehensively describing DNA methylation patterns spawns a diversity of DNA methylation profiling technologies that target its genomic distribution. These approaches have enabled the measurement of cytosine methylation from specific loci at restricted regions to single-base-pair resolution on a genome-scale level. In this review, we discuss the different DNA methylation analysis technologies primarily based on the initial treatments of DNA samples: bisulfite conversion, endonuclease digestion and affinity enrichment, involving methodology evolution, principles, applications, and their relative merits. This review may offer referable information for the selection of various platforms for genome-wide analysis of DNA methylation.
DNA 甲基化在真核生物中提供了一个关键的表观遗传调控层,对健康和疾病中的许多生物学过程都有重要影响。最初,人们提出 DNA 中胞嘧啶碱基的甲基化功能是一种“沉默”的表观遗传标记,并在几十年的时间里一直集中在基因的启动子区域。随着技术的改进和研究的积累,我们对 DNA 甲基化在各种基因组环境中的作用的理解不断扩展,包括基因体、重复序列和转录起始位点。对全面描述 DNA 甲基化模式的需求催生了多种针对其基因组分布的 DNA 甲基化分析技术。这些方法使我们能够从特定的受限区域的特定基因座测量胞嘧啶甲基化,甚至在基因组范围内达到单碱基分辨率。在这篇综述中,我们主要根据 DNA 样本的初始处理方法(亚硫酸氢盐转化、内切酶消化和亲和富集)讨论了不同的 DNA 甲基化分析技术,涉及方法学的演变、原理、应用及其相对优势。这篇综述可能为选择各种全基因组 DNA 甲基化分析平台提供参考信息。
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