CRISPR/Cas9 Off-targets: Computational Analysis of Causes, Prediction, Detection, and Overcoming Strategies

Roshan Kumar Roy, Ipsita Debashree, Sonal Srivastava, Narayan Rishi, Ashish Srivastava

Research output: Contribution to journalReview articlepeer-review

3 Scopus citations

Abstract

CRISPR/Cas9 technology is a highly flexible RNA-guided endonuclease (RGEN) based gene-editing tool that has transformed the field of genomics, gene therapy, and ge-nome/epigenome imaging. Its wide range of applications provides immense scope for understanding as well as manipulating genetic/epigenetic elements. However, the RGEN is prone to off-target mutagenesis that leads to deleterious effects. This review details the molecular and cel-lular mechanisms underlying the off-target activity, various available detection tools and prediction methodology ranging from sequencing to machine learning approaches, and the strategies to overcome/minimise off-targets. A coherent and concise method increasing target precision would prove indispensable to concrete manipulation and interpretation of genome editing results that can revolutionise therapeutics, including clarity in genome regulatory mechanisms during devel-opment.

Original languageEnglish (US)
Pages (from-to)119-132
Number of pages14
JournalCurrent Bioinformatics
Volume17
Issue number2
DOIs
StatePublished - Feb 2022
Externally publishedYes

Bibliographical note

Publisher Copyright:
© 2022 Bentham Science Publishers.

Keywords

  • chemical genetics
  • CRISPR/Cas9
  • drug mode of action
  • Genome-wide association study
  • pathogenesis
  • transcriptional regulation

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