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.
Keywords: N(6)-methyladenosine; Nanopore DRS; Next-generation sequencing; SMRT; m(6)A-mapping.
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