Abstract

Studies of chemical modifications on RNA have ushered in the field of epitranscriptomics. N6-Methyladenosine (m6A) is the most typical RNA modification and is indispensable for basic biological processes. This study presents a chemical pulldown method (m6A-ORL-Seq) for transcriptome-wide profiling of m6A. Moreover, chemical labeling results in a specific reverse transcription (RT) truncation signature. This study has identified four thousand high-confidence m6A sites at single-base resolution in the human transcriptome. Unlike previously reported methods based on m6A-antibody or m6A-sensitive enzymes, the antibody/enzyme-free chemical method provides a new perspective for single-base m6A detection at the transcriptome level.