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CRISPR/Cas Systems in Genome Editing: Methodologies and Tools for sgRNA Design, Off-Target Evaluation, and Strategies to Mitigate Off-Target Effects

文献类型： 外文期刊

作者： Manghwar, Hakim 1 ; Li, Bo 1 ; Ding, Xiao 1 ; Hussain, Amjad 1 ; Lindsey, Keith 3 ; Zhang, Xianlong 1 ; Jin, Shuangxia 1 ;

作者机构： 1.Huazhong Agr Univ, Natl Key Lab Crop Genet Improvement, Wuhan 430070, Hubei, Peoples R China

2.Xinjiang Acad Agr Sci, Inst Nucl & Biol Technol, Urumqi 830091, Xinjiang, Peoples R China

3.Univ Durham, Dept Biosci, Durham DH1 3LE, England

关键词： base editing; CRISPR; Cas systems; off-target effects; PAMs; sgRNA

期刊名称：ADVANCED SCIENCE （ 影响因子：16.806； 五年影响因子：17.835 ）

ISSN：

年卷期： 2020 年 7 卷 6 期

页码：

收录情况： SCI

摘要： Life sciences have been revolutionized by genome editing (GE) tools, including zinc finger nucleases, transcription activator-Like effector nucleases, and CRISPR (clustered regulatory interspaced short palindromic repeats)/Cas (CRISPR-associated) systems, which make the targeted modification of genomic DNA of all organisms possible. CRISPR/Cas systems are being widely used because of their accuracy, efficiency, and cost-effectiveness. Various classes of CRISPR/Cas systems have been developed, but their extensive use may be hindered by off-target effects. Efforts are being made to reduce the off-target effects of CRISPR/Cas9 by generating various CRISPR/Cas systems with high fidelity and accuracy. Several approaches have been applied to detect and evaluate the off-target effects. Here, the current GE tools, the off-target effects generated by GE technology, types of off-target effects, mechanisms of off-target effects, major concerns, and outcomes of off-target effects in plants and animals are summarized. The methods to detect off-target effects, tools for single-guide RNA (sgRNA) design, evaluation and prediction of off-target effects, and strategies to increase the on-target efficiency and mitigate the off-target impact on intended genome-editing outcomes are summarized.

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