基于CRISPR/Cas系统的基因编辑工具及其改进策略

Gene Editing Tools Based on CRISPR/Cas System and Their Improvement Strategies

CRISPR/Cas系统自发现以来持续推动着生命科学领域的进步。与此同时,CRISPR/Cas介导的基因编辑技术也在不断发展壮大。基于DSBs修复的CRISPR/Cas基因编辑技术、碱基编辑器和先导编辑器等新型基因编辑工具的开发为生物学基础研究铺平了道路。虽然这些工具为生物技术带来了革命性变化,但基因编辑效率偏低、产物纯度不高、脱靶效应频繁等问题也随之而来。不断开发精确、高效和安全的CRISPR/Cas基因编辑工具仍是当前和未来的生命科学研究热点。概述了CRISPR/Cas基因编辑工具的发展、构成及原理,总结了CRISPR/Cas基因编辑系统提升编辑效率、扩展编辑范围和降低脱靶效应的通用策略及不同CRISPR/Cas基因编辑工具的改进方法,并就CRISPR/Cas基因编辑工具未来的研究方向进行展望。

CRISPR/Cas system has continuously promoted the progress in the field of life science since its discovery.At the same time,the gene editing technology mediated by CRISPR/Cas is also developing and improving continuously.The development of new gene editing tools such as CRISPR/Cas gene editing technology based on DSBs repair,base editors and prime editors has paved the way for basic biological research.Although these tools have brought revolutionary changes to biotechnology,problems such as low efficiency of gene editing,low purity of products and frequent off⁃target effects began to emerge.The continuous development of accurate,efficient,and secure CRISPR/Cas editing tools remains a current and future research focus.Here,the development,composition and principle of CRISPR/Cas system based gene editing tools are briefly described.The general strategies for the CRISPR/Cas system to improve editing efficiency,expand editing scope and reduce off⁃target effects are systematically summarized,as well as the improvement methods of different CRISPR/Cas gene editing tools.In addition,the future research direction of CRISPR gene editing tools is discussed.