DNA methylation methods: Global DNA methylation and methylomic analyses

Methods. 2021 Mar:187:28-43. doi: 10.1016/j.ymeth.2020.10.002. Epub 2020 Oct 9.

Abstract

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.

Keywords: Affinity enrichment; Bisulfite conversion; DNA hydroxymethylation; DNA methylation; Endonuclease digestion; Microarray; Next-generation sequencing.

Publication types

  • Research Support, N.I.H., Extramural
  • Review

MeSH terms

  • Cytosine / metabolism
  • DNA Methylation*
  • Epigenomics / methods*
  • Genetic Loci
  • High-Throughput Nucleotide Sequencing / methods*
  • Humans
  • Whole Genome Sequencing / methods*

Substances

  • Cytosine