m6A-label-seq: A metabolic labeling protocol to detect transcriptome-wide mRNA N6-methyladenosine (m6A) at base resolution
Xiao Ou Shu, Jie Cao, Jianzhao Liu
Abstract
We describe here a metabolic labeling protocol for detecting cellular transcriptome-wide mRNA N6-methyladenosine (m6A) at base resolution. By feeding cells an analog of methionine, potential mRNA m6A forming sites are replaced with the N6-allyladenosine (a6A). A mild chemical iodination of a6A in RNA results in its opposite base misincorporation during RNA reverse transcription, and thus m6A locations could be precisely identified in the high-throughput sequencing. m6A-label-seq provides a strategy to label and identify cellular epitranscriptomic modification sites. For complete details on the use and execution of this profile, please refer to Shu et al., 2020.
Topics & Concepts
N6-MethyladenosineTranscriptomeRNAMessenger RNABiologyComputational biologyTranscription (linguistics)RNA-SeqBase pairGene expressionGeneGeneticsMethylationMethyltransferasePhilosophyLinguisticsRNA modifications and cancerCancer-related molecular mechanisms researchRNA and protein synthesis mechanisms