biostudies-literatureUnknown3(5)Shu XThe integration of RNA metabolic labelling by nucleoside analogues with high-throughput RNA sequencing has been harnessed to study RNA dynamics. The immunoprecipitation purification or chemical pulldown technique is generally required to enrich the analogue-labelled RNAs. Here we developed an a⁶A-seq method, which takes advantage of <i>N⁶-</i>allyladenosine (a⁶A) metabolic labelling on cellular mRNAs and profiles them in an immunoprecipitation-free and mutation-based manner. a⁶A plays a role as a chemical sequencing tag in that the iodination of a⁶A in mRNAs results in 1,<i>N</i> ⁶-cyclized adenosine (cyc-A), which induces base misincorporation during RNA reverse transcription, thus making a⁶A-labelled mRNAs detecFundamental research657-664https://www.ebi.ac.uk/biostudies/studies/S-EPMC11197751biostudies-literaturea⁶A-seq: <i>N</i> ⁶-allyladenosine-based cellular messenger RNA metabolic labelling and sequencing.PMC11197751Huang CLiu JLi TCao JShu Xfalsea⁶A-seq: <i>N</i> ⁶-allyladenosine-based cellular messenger RNA metabolic labelling and sequencing.The integration of RNA metabolic labelling by nucleoside analogues with high-throughput RNA sequencing has been harnessed to study RNA dynamics. The immunoprecipitation purification or chemical pulldown technique is generally required to enrich the analogue-labelled RNAs. Here we developed an a⁶A-seq method, which takes advantage of <i>N⁶-</i>allyladenosine (a⁶A) metabolic labelling on cellular mRNAs and profiles them in an immunoprecipitation-free and mutation-based manner. a⁶A plays a role as a chemical sequencing tag in that the iodination of a⁶A in mRNAs results in 1,<i>N</i> ⁶-cyclized adenosine (cyc-A), which induces base misincorporation during RNA reverse transcription, thus making a⁶A-labelled mRNAs detec2023-01-01T00:00:00Z2023 Sep2026-06-01T11:30:52.641Z2024-10-15T14:33:25.092ZS-EPMC111977513893329210.1016/j.fmre.2023.04.010