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

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submitted on 2024-06-02, 06:34 and posted on 2024-06-02, 06:39 authored by Maryam Saifaldeen, Dana E. Al-Ansari, Dindial Ramotar, Mustapha Aouida

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.

Other Information

Published in: Cells
License: https://creativecommons.org/licenses/by/4.0/
See article on publisher's website: https://dx.doi.org/10.3390/cells9112518

Funding

Qatar National Research Fund (QRLP11-G-2003014), Molecular and Cellular Biosciences.

Qatar National Research Fund (GSRA6-2-0603-19065), MSc-Genome wide screen in yeast to identify Beauvericin-Resistant and - Sensitive Target Genes”.

History

Language

  • English

Publisher

MDPI

Publication Year

  • 2020

License statement

This Item is licensed under the Creative Commons Attribution 4.0 International License.

Institution affiliated with

  • Hamad Bin Khalifa University
  • College of Health and Life Sciences - HBKU

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