Department of Life Sciences, King Fahd University of Petroleum and Minerals (KFUPM), Dhahran 31261, Saudi Arabia.
Department of Chemistry and Biochemistry, Texas Tech University, Lubbock, TX 79409, USA.
Cells. 2020 Jul 2;9(7):1608. doi: 10.3390/cells9071608.
Gene editing that makes target gene modification in the genome by deletion or addition has revolutionized the era of biomedicine. Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 emerged as a substantial tool due to its simplicity in use, less cost and extraordinary efficiency than the conventional gene-editing tools, including zinc finger nucleases (ZFNs) and Transcription activator-like effector nucleases (TALENs). However, potential off-target activities are crucial shortcomings in the CRISPR system. Numerous types of approaches have been developed to reduce off-target effects. Here, we review several latest approaches to reduce the off-target effects, including biased or unbiased off-target detection, cytosine or adenine base editors, prime editing, dCas9, Cas9 paired nickase, ribonucleoprotein (RNP) delivery and truncated gRNAs. This review article provides extensive information to cautiously interpret off-target effects to assist the basic and clinical applications in biomedicine.
基因编辑通过删除或添加来实现基因组中的靶基因修饰，从而彻底改变了生物医药时代。由于其使用简便、成本更低、效率比传统的基因编辑工具（如锌指核酸酶（ZFNs）和转录激活因子样效应核酸酶（TALENs））更高，因此，成簇规律间隔短回文重复序列（CRISPR）/Cas9 成为一种重要的工具。然而，潜在的脱靶活性是 CRISPR 系统的一个关键缺点。已经开发了许多种方法来降低脱靶效应。在这里，我们综述了几种最新的降低脱靶效应的方法，包括有偏或无偏的脱靶检测、胞嘧啶或腺嘌呤碱基编辑器、先导编辑、dCas9、Cas9 配对切口酶、核糖核蛋白（RNP）递送和截断 gRNA。这篇综述文章提供了广泛的信息，以谨慎解释脱靶效应，以协助基础和临床医学中的应用。
