Bioinformatic Tools in CRISPR/Cas Platform

Aftab Ahmad, Sidra Ashraf, Humera Naz Majeed, Sabin Aslam, Muhammad Aamir Aslam, Muhammad Salman Mubarik, Nayla Munawar

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

CRISPR/Cas has emerged as a game-changing technology for genome editing with widespread applications ranging from human therapeutics to engineering bacterial genomes for beneficial purposes to editing plant genomes for agricultural purposes. Successful genome editing through CRISPR/Cas relies on two components: an appropriate Cas endonuclease and a 20-base-pair (bp), singleguide RNA (sgRNA). CRISPR/Cas is currently favored as a genome editing technique due to its simple design rules and efficient editing capabilities that do not necessarily involve adding any foreign DNA at the target site. Cas endonucleases can be programmed to target any site in the genome by changing the gRNA sequence, highlighting the importance of gRNA design for increased specificity and efficiency, and reduced off-targeting in CRISPR/Cas genome editing. The rapid rise in CRISPR/Cas genome editing and associated applications has led to the development of numerous computational tools for effective sgRNA design. In this chapter, we discuss the essentials of gRNA design and provide an overview of the design process. In addition to summarizing factors which affect gRNA specificity and CRISPR cleavage efficiency, we discuss predictions of target efficiency and off-target detection algorithms. Finally, we describe the application-specific (knockout, activation, repression, base editing, and RNA editing) requirements of gRNA design and different tools to facilitate gRNA design.

Original languageEnglish
Title of host publicationThe CRISPR/Cas Tool Kit for Genome Editing
PublisherSpringer Nature
Pages53-111
Number of pages59
ISBN (Electronic)9789811663055
ISBN (Print)9789811663048
DOIs
Publication statusPublished - Jan 1 2022

Keywords

  • Bioinformatic tools
  • CRISPR/Cas
  • Design tools
  • Off-targeting
  • On-targeting efficiency
  • Specificity of CRISPR/Cas

ASJC Scopus subject areas

  • General Medicine
  • General Health Professions

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