Transcriptome-wide identification of A-to-I RNA editing sites using ICE-seq

Methods. 2019 Mar 1:156:66-78. doi: 10.1016/j.ymeth.2018.12.007. Epub 2018 Dec 20.

Abstract

In A-to-I RNA editing, adenosine is converted to inosine in double-stranded regions of RNAs. Inosine, an abundant epitranscriptomic mark, contributes to a wide range of biological processes by regulating gene expression post-transcriptionally. To understand the effect of A-to-I RNA editing on regulation of the epitranscriptome, accurate mapping of inosines is necessary. To this end, we established a biochemical method called inosine chemical erasing sequencing (ICE-seq) that enables unbiased and reliable identification of A-to-I RNA editing sites throughout the transcriptome. Here, we describe our updated protocol for ICE-seq in the human transcriptome.

Keywords: A-to-I RNA editing; High-throughput sequencing; ICE-seq.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine / chemistry
  • Adenosine / metabolism
  • Animals
  • DNA, Complementary / chemistry
  • DNA, Complementary / genetics
  • DNA, Complementary / metabolism
  • Ethanol / chemistry
  • Ethylamines / chemistry
  • High-Throughput Nucleotide Sequencing / methods*
  • Humans
  • Inosine / chemistry
  • Inosine / metabolism
  • Molecular Sequence Annotation / methods*
  • Nitriles / chemistry
  • Poly A / chemistry
  • Poly A / genetics
  • Poly A / metabolism
  • RNA Editing*
  • RNA, Double-Stranded / chemistry
  • RNA, Double-Stranded / genetics
  • RNA, Double-Stranded / metabolism
  • RNA, Messenger / chemistry*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Transcriptome*

Substances

  • DNA, Complementary
  • Ethylamines
  • Nitriles
  • RNA, Double-Stranded
  • RNA, Messenger
  • Poly A
  • Ethanol
  • Inosine
  • Adenosine
  • triethylamine