Zhang Gui Shan, Yang Yong, Zhang Ling Min, Dai Xian Hua
School of Electronics and Information Technology, Sun Yat-sen University, Guangzhou 510006, China.
Yi Chuan. 2018 Sep 20;40(9):704-723. doi: 10.16288/j.yczz.18-135.
The third generation of the CRISPR/Cas9-mediated genome fixed-point editing technology has been widely used in the field of gene editing and gene expression regulation. How to improve the on-target efficiency and specificity of this system, as well as reduce its off-target effects are always the bottleneck in its development. Machine learning provides novel methods to the problems of the CRISPR/Cas9 system, and CRISPR/Cas9-based machine learning has recently become a very hot research topic. In this review, we firstly outline the mechanism of the CRISPR/Cas9 system. Subsequently, we elaborate the current issues of CRISPR/Cas9, including low efficiency and potential off-target effects, and sequence-recognizing limitation from protospacer adjacent motif (PAM). Finally, we summarize the applications of methods within the machine learning framework for optimizing the CRISPR/Cas9 system, such as optimized single-guide RNA (sgRNA) design, CRISPR/Cas9 cleavage efficiency prediction, off-target effects evaluation, gene knock-out as well as high-throughput functional genetic screening and prospects for development.
第三代CRISPR/Cas9介导的基因组定点编辑技术已在基因编辑和基因表达调控领域得到广泛应用。如何提高该系统的靶向效率和特异性,以及降低其脱靶效应,一直是其发展的瓶颈。机器学习为CRISPR/Cas9系统的问题提供了新方法,基于CRISPR/Cas9的机器学习最近成为一个非常热门的研究课题。在这篇综述中,我们首先概述了CRISPR/Cas9系统的机制。随后,我们阐述了CRISPR/Cas9当前存在的问题,包括效率低下和潜在的脱靶效应,以及原间隔相邻基序(PAM)的序列识别限制。最后,我们总结了机器学习框架内用于优化CRISPR/Cas9系统的方法的应用,如优化的单向导RNA(sgRNA)设计、CRISPR/Cas9切割效率预测、脱靶效应评估、基因敲除以及高通量功能基因筛选和发展前景。