Strategies for DNA methylation analysis in developmental studies

Abstract

Developmental processes in eukaryotes are highly dependent on DNA methylation. 5-methylcytosine (m5C) is the most prevalent and best understood DNA modification implicated in maintenance of genomic integrity and function across species. Although m5C occurs almost exclusively in symmetrical CpG context in vertebrates, additional asymmetrical distribution in CpHpG and CpHpH sites has been observed in plants and embryonic stem cells. To this end, accurate and reproducible methodology for full analysis of the DNA methylome is highly demanded. Fortunately, a variety of methods enable quantitative DNA methylation mapping at a single-base resolution and in a large scale. Here, we provide a critical overview of methods applied primarily to m5C detection with particular emphasis on technical improvements of the classical bisulfite-conversion protocol. We further describe strategies in combination with emerging technologies that allow acquisition of highly reliable data for developmental studies. © 2011 The Authors. Journal compilation © 2011 Japanese Society of Developmental Biologists.