CRISPR/Cas9 Guide RNA Design Rules for Predicting Activity

Methods Mol Biol. 2020:2115:351-364. doi: 10.1007/978-1-0716-0290-4_19.

Abstract

A critical stage in performing gene editing experiments using the CRISPR/Cas9 system is the design of guide RNA (gRNA). In this chapter, we conduct a review of the current gRNA design rules for maximizing on-target Cas9 activity while minimizing off-target activity. In addition, we present some of the currently available computational tools for gRNA activity prediction and assay design.

Keywords: CRISPR; Cas9; Genome editing; Machine learning; sgRNA.

Publication types

  • Review

MeSH terms

  • Animals
  • CRISPR-Associated Protein 9 / genetics
  • CRISPR-Cas Systems*
  • Clustered Regularly Interspaced Short Palindromic Repeats
  • Gene Editing / methods
  • Genomics / methods
  • Humans
  • Models, Genetic
  • RNA, Guide, CRISPR-Cas Systems / genetics*

Substances

  • RNA, Guide, CRISPR-Cas Systems
  • CRISPR-Associated Protein 9