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

Over 160 RNA modifications have been identified, including N⁷-methylguanine (m⁷G), N⁶-methyladenosine (m⁶A), and 5-methylcytosine (m⁵C). These modifications play key roles in regulating the fate of RNA. In eukaryotes, m⁶A is the most abundant mRNA modification, accounting for over 80% of all RNA methylation modifications. Highly dynamic m⁶A modification may exert important effects on organismal reproduction and development. Significant advances in understanding the mechanism of m⁶A 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 m⁶A RNA modifications, emphasizing technological advances in detecting m⁶A modification. In particular, we discuss the challenge of generating accurate dynamic single-base resolution m⁶A 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 m⁶A modification.