CRISPR FokI Dead Cas9 System: Principles and Applications in Genome Engineering

Maryam Saifaldeen, Dana E. Al-Ansari, Dindial Ramotar, Mustapha Aouida

Research output: Contribution to journalReview articlepeer-review

26 Citations (Scopus)

Abstract

The identification of the robust clustered regularly interspersed short palindromic repeats (CRISPR) associated endonuclease (Cas9) system gene-editing tool has opened up a wide range of potential therapeutic applications that were restricted by more complex tools, including zinc finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs). Nevertheless, the high frequency of CRISPR system off-target activity still limits its applications, and, thus, advanced strategies for highly specific CRISPR/Cas9-mediated genome editing are continuously under development including CRISPR-FokI dead Cas9 (fdCas9). fdCas9 system is derived from linking a FokI endonuclease catalytic domain to an inactive Cas9 protein and requires a pair of guide sgRNAs that bind to the sense and antisense strands of the DNA in a protospacer adjacent motif (PAM)-out orientation, with a defined spacer sequence range around the target site. The dimerization of FokI domains generates DNA double-strand breaks, which activates the DNA repair machinery and results in genomic edit. So far, all the engineered fdCas9 variants have shown promising gene-editing activities in human cells when compared to other platforms. Herein, we review the advantages of all published variants of fdCas9 and their current applications in genome engineering.

Original languageEnglish
Article number2513
JournalCells
Volume9
Issue number11
DOIs
Publication statusPublished - 21 Nov 2020

Keywords

  • CRISPR
  • FokI endonuclease
  • FokI-dCas9
  • RNA-guided FokI nuclease
  • dead Cas9
  • dual guide RNA
  • fdCas9 variants
  • genome engineering
  • spacer sequence

Fingerprint

Dive into the research topics of 'CRISPR FokI Dead Cas9 System: Principles and Applications in Genome Engineering'. Together they form a unique fingerprint.

Cite this