Transcriptome-wide profiling of N⁶-methyladenosine via a selective chemical labeling method

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.