Detection of N6-methyladenosine in SARS-CoV-2 RNA by methylated RNA immunoprecipitation sequencing

STAR Protoc. 2022 Mar 18;3(1):101067. doi: 10.1016/j.xpro.2021.101067. Epub 2021 Dec 8.

Abstract

N 6 -methylation of adenosine (m6A) is the most abundant internal mRNA modification and is an important post-transcriptional regulator of gene expression. Here, we describe a protocol for methylated RNA immunoprecipitation sequencing (MeRIP-Seq) to detect and quantify m6A modifications in severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA. The protocol is optimized for low viral RNA levels and is readily adaptable for other applications. For complete details on the use and execution of this protocol, please refer to Li et al. (2021).

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Adenosine / analogs & derivatives*
  • Adenosine / analysis
  • Adenosine / genetics
  • Animals
  • COVID-19 / genetics
  • Caco-2 Cells
  • Chlorocebus aethiops
  • Gene Expression / genetics
  • Gene Expression Regulation / genetics
  • Genetic Techniques
  • HEK293 Cells
  • Humans
  • Immunoprecipitation / methods*
  • Methylation
  • RNA / chemistry
  • RNA / genetics
  • RNA Processing, Post-Transcriptional
  • RNA, Viral / metabolism
  • SARS-CoV-2 / genetics
  • SARS-CoV-2 / pathogenicity
  • Sequence Analysis, RNA / methods*
  • Vero Cells

Substances

  • RNA, Viral
  • RNA
  • N-methyladenosine
  • Adenosine