2015 August ; 12(8): 767–772 | Bastian Linder, Anya V. Grozhik, Anthony O. Olarerin-George, Cem Meydan, Christopher E. Mason, Samie R. Jaffrey
This study introduces a novel method called "m6A individual-nucleotide resolution crosslinking and immunoprecipitation" (miCLIP) to map m6A and m6Am residues at single-nucleotide resolution in human and mouse mRNA. The method involves crosslinking anti-m6A antibodies to RNA using UV light, followed by reverse transcription, which induces specific mutations or truncations in the cDNA that indicate the presence of the bound protein. The authors tested different anti-m6A antibodies and found that the Abcam and SySy polyclonal antibodies produced consistent patterns of mutations and truncations, respectively. Using these antibodies, they mapped m6A and m6Am residues throughout the transcriptome, identifying snoRNAs as a novel class of m6A-containing non-coding RNAs. The miCLIP method provides a highly specific and sensitive way to identify m6A residues, allowing for detailed functional studies of m6A in transcripts.This study introduces a novel method called "m6A individual-nucleotide resolution crosslinking and immunoprecipitation" (miCLIP) to map m6A and m6Am residues at single-nucleotide resolution in human and mouse mRNA. The method involves crosslinking anti-m6A antibodies to RNA using UV light, followed by reverse transcription, which induces specific mutations or truncations in the cDNA that indicate the presence of the bound protein. The authors tested different anti-m6A antibodies and found that the Abcam and SySy polyclonal antibodies produced consistent patterns of mutations and truncations, respectively. Using these antibodies, they mapped m6A and m6Am residues throughout the transcriptome, identifying snoRNAs as a novel class of m6A-containing non-coding RNAs. The miCLIP method provides a highly specific and sensitive way to identify m6A residues, allowing for detailed functional studies of m6A in transcripts.