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

Autor:	Mustapha Aouida, Maryam Saifaldeen, Dindial Ramotar, Dana E Al-Ansari
Rok vydání:	2020
Předmět:	spacer sequence FokI-dCas9 Review Computational biology RNA-guided FokI nuclease Genome engineering Genome editing Humans CRISPR Deoxyribonucleases Type II Site-Specific lcsh:QH301-705.5 fdCas9 variants Gene Editing Transcription activator-like effector nuclease Tissue Engineering biology Cas9 Genomics General Medicine Zinc finger nuclease FokI Protospacer adjacent motif dual guide RNA lcsh:Biology (General) biology.protein FokI endonuclease CRISPR-Cas Systems genome engineering dead Cas9
Zdroj:	Cells, Vol 9, Iss 2518, p 2518 (2020) Cells
ISSN:	2073-4409
DOI:	10.3390/cells9112518
Popis:	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.
Databáze:	OpenAIRE
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