摘要
酿酒酵母是常用的工业生产菌株,改造酿酒酵母以提高其生产性能极为重要。然而,传统的改造方法存在步骤繁琐、周期长等问题。CRISPR/Cas9系统是在细菌和古细菌发现的自适应防御系统。目前CRISPR/Cas9系统已经成为基因组编辑的有力工具,能够同时改造酿酒酵母的多个基因。本文综述CRISPR/Cas9系统各组分的作用和系统的构建,介绍sgRNA靶序列的设计原则和代表性设计工具,总结对酿酒酵母进行多基因编辑的策略,以及CRISPR/Cas9系统存在脱靶和编辑效率低的问题和应对展望,为更好地应用CRISPR/Cas9系统改造酿酒酵母提供参考。
Saccharomyces cerevisiae is one of widely used industrial strains,it is extremely important to increase its performance via modifying S.cerevisiae;however,there are cumbersome steps and long-cycle by traditional modifying method.CRISPR/Cas9 system is an adaptive defense system found in bacteria and archaea.At present,the CRISPR/Cas9 system has become a powerful tool for genome editing,which can simultaneously modify multiple genes of S.cerevisiae.This review provided a brief overview of the function and construction of CRISPR/Cas9 system in S.cerevisiae gene editing,introduced the design rules and representative design tools of sgRNA targeting sequences,as well as summarized the multiple-gene editing strategy.Moreover,the review discussed the challenges in application of CRISPR/Cas9 system,including off target and low editing efficiency,and their control measures,which may provide the references for better applying CRISPR/Cas9 system modifying S.cerevisiae.
作者
陈小玲
廖东庆
黄尚飞
陈英
芦志龙
陈东
CHEN Xiao-ling;LIAO Dong-qing;HUANG Shang-fei;CHEN Ying;LU Zhi-long;CHEN Dong(Guangxi Academy of Sciences,National key Laboratory of Non-food Biomass Energy Technology,State Key Laboratory of Non-food Biomass and Enzyme Technology,Nanning 530007;Guangxi Light Industry Science and Technology Research Institute Co.,Ltd.,Nanning 530031)
出处
《生物技术通报》
CAS
CSCD
北大核心
2023年第8期148-158,共11页
Biotechnology Bulletin
基金
广西自然科学基金项目(2020GXNSFBA297070,2023GXNSFAA026462)。
