MA RNA Immunoprecipitation Followed by High-Throughput Sequencing to Map N-Methyladenosine

Overview

Journal Methods Mol Biol

Specialty Molecular Biology

Date 2021 Oct 25

PMID 34694619

Citations 5

Authors

Devi Prasad Bhattarai

Francesca Aguilo

Affiliations

Soon will be listed here.

Abstract

N-methyladenosine (mA) is the most abundant internal modification on messenger RNAs (mRNAs) and long noncoding RNAs (lncRNAs) in eukaryotes. It influences gene expression by regulating RNA processing, nuclear export, mRNA decay, and translation. Hence, mA controls fundamental cellular processes, and dysregulated deposition of mA has been acknowledged to play a role in a broad range of human diseases, including cancer. mA RNA immunoprecipitation followed by high-throughput sequencing (MeRIP-seq or mA-seq) is a powerful technique to map mA in a transcriptome-wide level. After immunoprecipitation of fragmented polyadenylated (poly(A)) rich RNA by using specific anti-mA antibodies, both the immunoprecipitated RNA fragments together with the input control are subjected to massively parallel sequencing. The generation of such comprehensive methylation profiles of signal enrichment relative to input control is necessary in order to better comprehend the pathogenesis behind aberrant mA deposition.

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