Advances in the profiling of N6-methyladenosine (m6A) modifications

Over 160 RNA modifications have been identified, including N-7-methylguanine (m(7)G), N-6-methyladenosine (m(6)A), and 5-methylcytosine (m(5)C). These modifications play key roles in regulating the fate of RNA. In eukaryotes, m(6)A is the most abundant mRNA modification, accounting for over 80% of all RNA methylation modifications. Highly dynamic m(6)A modification may exert important effects on organismal reproduction and development. Significant advances in understanding the mechanism of m(6)A modification have been made using immunoprecipitation, chemical labeling, and site-directed mutagenesis, combined with next-generation sequencing. Single-molecule real-time and nanopore direct RNA sequencing (DRS) approaches provide additional ways to study RNA modifications at the cellular level. In this review, we explore the technical history of identifying m(6)A RNA modifications, emphasizing technological advances in detecting m(6)A modification. In particular, we discuss the challenge of generating accurate dynamic single-base resolution m(6)A maps and also strategies for improving detection specificity. Finally, we outline a roadmap for future research in this area, focusing on the application of RNA epigenetic modification, represented by m(6)A modification.