Our official English website, www.x-mol.net, welcomes your
feedback! (Note: you will need to create a separate account there.)
Cas9 Cuts and Consequences; Detecting, Predicting, and Mitigating CRISPR/Cas9 On- and Off-Target Damage: Techniques for Detecting, Predicting, and Mitigating the On- and off-target Effects of Cas9 Editing.
BioEssays ( IF 3.2 ) Pub Date : 2020-07-09 , DOI: 10.1002/bies.202000047 Anthony Newman 1 , Lora Starrs 1 , Gaetan Burgio 1
BioEssays ( IF 3.2 ) Pub Date : 2020-07-09 , DOI: 10.1002/bies.202000047 Anthony Newman 1 , Lora Starrs 1 , Gaetan Burgio 1
Affiliation
|
Large deletions and genomic re‐arrangements are increasingly recognized as common products of double‐strand break repair at Clustered Regularly Interspaced, Short Palindromic Repeats ‐ CRISPR associated protein 9 (CRISPR/Cas9) on‐target sites. Together with well‐known off‐target editing products from Cas9 target misrecognition, these are important limitations, that need to be addressed. Rigorous assessment of Cas9‐editing is necessary to ensure validity of observed phenotypes in Cas9‐edited cell‐lines and model organisms. Here the mechanisms of Cas9 specificity, and strategies to assess and mitigate unwanted effects of Cas9 editing are reviewed; covering guide‐RNA design, RNA modifications, Cas9 modifications, control of Cas9 activity; computational prediction for off‐targets, and experimental methods for detecting Cas9 cleavage. Although recognition of the prevalence of on‐ and off‐target effects of Cas9 editing has increased in recent years, broader uptake across the gene editing community will be important in determining the specificity of Cas9 across diverse applications and organisms.
中文翻译：
Cas9 削减和后果；检测、预测和减轻 CRISPR/Cas9 靶标和脱靶损伤：检测、预测和减轻 Cas9 编辑的靶标和脱靶效应的技术。
大缺失和基因组重排越来越被认为是成簇规则间隔短回文重复 CRISPR 相关蛋白 9 (CRISPR/Cas9) 靶位点双链断裂修复的常见产物。加上众所周知的 Cas9 目标误识别导致的脱靶编辑产品，这些都是需要解决的重要限制。对 Cas9 编辑进行严格评估对于确保 Cas9 编辑细胞系和模型生物中观察到的表型的有效性是必要的。本文回顾了 Cas9 特异性的机制，以及评估和减轻 Cas9 编辑的不良影响的策略；涵盖引导RNA设计、RNA修饰、Cas9修饰、Cas9活性控制；脱靶的计算预测以及检测 Cas9 裂解的实验方法。尽管近年来人们对 Cas9 编辑的脱靶效应和脱靶效应的普遍认识有所增加，但基因编辑界更广泛的采用对于确定 Cas9 在不同应用和生物体中的特异性非常重要。
更新日期：2020-08-26
中文翻译：
Cas9 削减和后果；检测、预测和减轻 CRISPR/Cas9 靶标和脱靶损伤：检测、预测和减轻 Cas9 编辑的靶标和脱靶效应的技术。
大缺失和基因组重排越来越被认为是成簇规则间隔短回文重复 CRISPR 相关蛋白 9 (CRISPR/Cas9) 靶位点双链断裂修复的常见产物。加上众所周知的 Cas9 目标误识别导致的脱靶编辑产品，这些都是需要解决的重要限制。对 Cas9 编辑进行严格评估对于确保 Cas9 编辑细胞系和模型生物中观察到的表型的有效性是必要的。本文回顾了 Cas9 特异性的机制，以及评估和减轻 Cas9 编辑的不良影响的策略；涵盖引导RNA设计、RNA修饰、Cas9修饰、Cas9活性控制；脱靶的计算预测以及检测 Cas9 裂解的实验方法。尽管近年来人们对 Cas9 编辑的脱靶效应和脱靶效应的普遍认识有所增加，但基因编辑界更广泛的采用对于确定 Cas9 在不同应用和生物体中的特异性非常重要。
京公网安备 11010802027423号