Discovering a-to-i rna editing through chemical methodology “ICE-SEQ”

Masayuki Sakurai, Shunpei Okada, Hiroki Ueda, Yuxi Yang

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

Abstract

RNA editing of adenosines to inosines contributes to a wide range of biological processes by regulating gene expression post-transcriptionally. To understand the effect, accurate mapping of inosines is necessary. The most conventional method to identify an editing site is to compare the cDNA sequence with its corresponding genomic sequence. However, this method has a high false discovery rate because guanosine signals, due to experimental errors or noise in the obtained sequences, contaminate genuine inosine signals detected as guanosine. To ensure high accuracy, we developed the Inosine Chemical Erasing (ICE) method to accurately and biochemically identify inosines in RNA strands utilizing inosine cyanoethylation and reverse transcription-PCR. Furthermore, we applied this technique to next-generation sequencing technology, called ICE-seq, to conduct an unbiased genome-wide screening of A-to-I editing sites in the transcriptome.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages113-148
Number of pages36
DOIs
Publication statusPublished - 2021

Publication series

NameMethods in Molecular Biology
Volume2181
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • A-to-I RNA editing
  • ICE-seq
  • Inosine cyanoethylation

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