Safari Fatemeh, Farajnia Safar, Ghasemi Younes, Zarghami Nosratollah
Department of Medical Biotechnology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran.
Student Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran.
Curr Pharm Biotechnol. 2017;18(13):1038-1054. doi: 10.2174/1389201019666180209120533.
RNA-guided endonuclease as a versatile genome editing technology opened new windows in various fields of biology. The simplicity of this revolutionary technique provides a promising future for its application in a broad range of approaches from functional annotation of genes to diseases, to genetic manipulation and gene therapy. Besides the site-specific activity of Cas9 endonuclease, the unintended cleavage known as off-target effect is still a major challenge for this genome editing technique.
Various strategies have been developed to resolve this bottleneck including development of new softwares for designing optimized guide RNA (gRNA), engineering Cas9 enzyme, improvement in off-target detection assays, etc. Results: This review dedicated to discuss on methods that have been used for optimizing Cas9, specificity with the aim of improving this technology for therapeutic applications.
In addition, the applications and novel breakthroughs in the field of CRISPR technology will be described.
RNA引导的核酸内切酶作为一种多功能的基因组编辑技术，为生物学的各个领域打开了新的窗口。这项革命性技术的简单性为其在从基因功能注释到疾病、基因操作和基因治疗等广泛方法中的应用提供了广阔的前景。除了Cas9核酸内切酶的位点特异性活性外，被称为脱靶效应的意外切割仍然是这种基因组编辑技术的一个主要挑战。
已经开发了各种策略来解决这一瓶颈，包括开发用于设计优化引导RNA（gRNA）的新软件、改造Cas9酶、改进脱靶检测方法等。
本综述致力于讨论用于优化Cas9的方法、特异性，旨在改进该技术用于治疗应用。
此外，还将描述CRISPR技术领域的应用和新突破。
