a6A-seq: N6-allyladenosine-based cellular messenger RNA metabolic labelling and sequencing

In this work, we applied a6A to label and sequence cellular mRNA in an immunoprecipitation-free and mutation-based manner, termed a6A-seq. a6A-seq was utilized to study the change of cellular gene expression profile under a methionine-free stressed condition. Compared with regular RNA-seq, a6A-seq could more sensitively detect the level change of newly synthesized mRNAs. Overall design: Profiles of a6A-labeled mRNA in HeLa cells (methionine-deficient and methionine-sufficient treatment) were generated by high-throughput sequencing, in duplicate, using HiSeq X Ten.