High-Resolution Mapping of N 6-Methyladenosine Using m6A Crosslinking Immunoprecipitation Sequencing (m6A-CLIP-Seq)

Methods Mol Biol. 2019:1870:69-79. doi: 10.1007/978-1-4939-8808-2_5.

Abstract

N 6-Methyladenosine, an abundant chemical modification in mRNA, plays crucial roles in regulating gene expression and biological processes. Research on m6A and its functions has progressed rapidly in the past few years, aided substantially by advances in high-throughput sequencing-based methods to profile m6A along the transcriptome. We present here a protocol for m6A crosslinking immunoprecipitation sequencing (m6A-CLIP-seq), which profiles m6A on mRNA at high resolution from as little as 1 μg of poly(A)-selected mRNA.

Keywords: Affinity purification; Methylome; N 6-Methyladenosine; Sequencing; Transcriptome.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine / metabolism*
  • HeLa Cells
  • High-Throughput Nucleotide Sequencing / methods*
  • Humans
  • Immunoprecipitation / methods
  • Methylation
  • RNA / genetics*
  • RNA / isolation & purification
  • RNA / metabolism*

Substances

  • RNA
  • Adenosine