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Evaluating and Enhancing Target Specificity of Gene-Editing Nucleases and Deaminases

Cited 31 time in Web of Science Cited 38 time in Scopus
Authors
Kim, Daesik; Luk, Kevin; Wolfe, Scot A.; Kim, Jin-Soo
Issue Date
2019-03
Citation
Annual Review of Biochemistry, Vol.88, pp.191-220
Keywords
CRISPR/Cas9off-targetgene editingbase editorsCas12aCpf1
Abstract
Programmable nucleases and deaminases, which include zinc-finger nucleases, transcription activator-like effector nucleases, CRISPR RNA-guided nucleases, and RNA-guided base editors, are now widely employed for the targeted modification of genomes in cells and organisms. These gene-editing tools hold tremendous promise for therapeutic applications. Importantly, these nucleases and deaminases may display off-target activity through the recognition of near-cognate DNA sequences to their target sites, resulting in collateral damage to the genome in the form of local mutagenesis or genomic rearrangements. For therapeutic genome-editing applications with these classes of programmable enzymes, it is essential to measure and limit genome-wide off-target activity. Herein, we discuss the key determinants of off-target activity for these systems. We describe various cell-based and cell-free methods for identifying genome-wide off-target sites and diverse strategies that have been developed for reducing the off-target activity of programmable gene-editing enzymes.
ISSN
0066-4154
URI
https://hdl.handle.net/10371/171744
DOI
https://doi.org/10.1146/annurev-biochem-013118-111730
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