Affiliation:
1. Department of Chemistry The University of Chicago Chicago IL USA
2. Institute for Biophysical Dynamics The University of Chicago Chicago IL USA
3. Program in Cellular and Molecular Medicine Boston Children's Hospital Boston MA USA
Abstract
AbstractN6‐methyladenosine (m6A) is the most abundant internal modification in mammalian messenger RNA (mRNA), constituting 0.1 %–0.4 % of total adenosine residues in the transcriptome. m6A regulates mRNA stability and translation, pre‐mRNA splicing, miRNA biogenesis, lncRNA binding, and many other physiological and pathological processes. While the majority of m6As occur in a consensus motif of DRm6ACH (D=A/G/U, R=A/G, H=U/A/C), the presence of such a motif does not guarantee methylation. Different RNA copies transcribed from the same gene may be methylated to varying levels. Within a single transcript, m6As are not evenly distributed, showing an enrichment in long internal and terminal exons. These characteristics of m6A deposition call for sequencing methods that not only pinpoint m6A sites at base resolution, but also quantitate the abundance of methylation across different RNA copies. In this review, we summarize existing m6A profiling methods, with an emphasis on next generation sequencing‐(NGS−)based, site‐specific, and quantitative methods, as well as several emerging single‐cell methods.