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Strategies for High-Efficiency Mutation Using the CRISPR/Cas System

Clustered regularly interspaced short palindromic repeats (CRISPR)-associated systems have revolutionized traditional gene-editing tools and are a significant tool for ameliorating gene defects. Characterized by high target specificity, extraordinary efficiency, and cost-effectiveness, CRISPR/Cas sy...

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Autores principales: Feng, Shuying, Wang, Zilong, Li, Aifang, Xie, Xin, Liu, Junjie, Li, Shuxuan, Li, Yalan, Wang, Baiyan, Hu, Lina, Yang, Lianhe, Guo, Tao
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8860194/
https://www.ncbi.nlm.nih.gov/pubmed/35198566
http://dx.doi.org/10.3389/fcell.2021.803252
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author Feng, Shuying
Wang, Zilong
Li, Aifang
Xie, Xin
Liu, Junjie
Li, Shuxuan
Li, Yalan
Wang, Baiyan
Hu, Lina
Yang, Lianhe
Guo, Tao
author_facet Feng, Shuying
Wang, Zilong
Li, Aifang
Xie, Xin
Liu, Junjie
Li, Shuxuan
Li, Yalan
Wang, Baiyan
Hu, Lina
Yang, Lianhe
Guo, Tao
author_sort Feng, Shuying
collection PubMed
description Clustered regularly interspaced short palindromic repeats (CRISPR)-associated systems have revolutionized traditional gene-editing tools and are a significant tool for ameliorating gene defects. Characterized by high target specificity, extraordinary efficiency, and cost-effectiveness, CRISPR/Cas systems have displayed tremendous potential for genetic manipulation in almost any organism and cell type. Despite their numerous advantages, however, CRISPR/Cas systems have some inherent limitations, such as off-target effects, unsatisfactory efficiency of delivery, and unwanted adverse effects, thereby resulting in a desire to explore approaches to address these issues. Strategies for improving the efficiency of CRISPR/Cas-induced mutations, such as reducing off-target effects, improving the design and modification of sgRNA, optimizing the editing time and the temperature, choice of delivery system, and enrichment of sgRNA, are comprehensively described in this review. Additionally, several newly emerging approaches, including the use of Cas variants, anti-CRISPR proteins, and mutant enrichment, are discussed in detail. Furthermore, the authors provide a deep analysis of the current challenges in the utilization of CRISPR/Cas systems and the future applications of CRISPR/Cas systems in various scenarios. This review not only serves as a reference for improving the maturity of CRISPR/Cas systems but also supplies practical guidance for expanding the applicability of this technology.
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spelling pubmed-88601942022-02-22 Strategies for High-Efficiency Mutation Using the CRISPR/Cas System Feng, Shuying Wang, Zilong Li, Aifang Xie, Xin Liu, Junjie Li, Shuxuan Li, Yalan Wang, Baiyan Hu, Lina Yang, Lianhe Guo, Tao Front Cell Dev Biol Cell and Developmental Biology Clustered regularly interspaced short palindromic repeats (CRISPR)-associated systems have revolutionized traditional gene-editing tools and are a significant tool for ameliorating gene defects. Characterized by high target specificity, extraordinary efficiency, and cost-effectiveness, CRISPR/Cas systems have displayed tremendous potential for genetic manipulation in almost any organism and cell type. Despite their numerous advantages, however, CRISPR/Cas systems have some inherent limitations, such as off-target effects, unsatisfactory efficiency of delivery, and unwanted adverse effects, thereby resulting in a desire to explore approaches to address these issues. Strategies for improving the efficiency of CRISPR/Cas-induced mutations, such as reducing off-target effects, improving the design and modification of sgRNA, optimizing the editing time and the temperature, choice of delivery system, and enrichment of sgRNA, are comprehensively described in this review. Additionally, several newly emerging approaches, including the use of Cas variants, anti-CRISPR proteins, and mutant enrichment, are discussed in detail. Furthermore, the authors provide a deep analysis of the current challenges in the utilization of CRISPR/Cas systems and the future applications of CRISPR/Cas systems in various scenarios. This review not only serves as a reference for improving the maturity of CRISPR/Cas systems but also supplies practical guidance for expanding the applicability of this technology. Frontiers Media S.A. 2022-02-07 /pmc/articles/PMC8860194/ /pubmed/35198566 http://dx.doi.org/10.3389/fcell.2021.803252 Text en Copyright © 2022 Feng, Wang, Li, Xie, Liu, Li, Li, Wang, Hu, Yang and Guo. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Cell and Developmental Biology
Feng, Shuying
Wang, Zilong
Li, Aifang
Xie, Xin
Liu, Junjie
Li, Shuxuan
Li, Yalan
Wang, Baiyan
Hu, Lina
Yang, Lianhe
Guo, Tao
Strategies for High-Efficiency Mutation Using the CRISPR/Cas System
title Strategies for High-Efficiency Mutation Using the CRISPR/Cas System
title_full Strategies for High-Efficiency Mutation Using the CRISPR/Cas System
title_fullStr Strategies for High-Efficiency Mutation Using the CRISPR/Cas System
title_full_unstemmed Strategies for High-Efficiency Mutation Using the CRISPR/Cas System
title_short Strategies for High-Efficiency Mutation Using the CRISPR/Cas System
title_sort strategies for high-efficiency mutation using the crispr/cas system
topic Cell and Developmental Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8860194/
https://www.ncbi.nlm.nih.gov/pubmed/35198566
http://dx.doi.org/10.3389/fcell.2021.803252
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