利用CRISPR/Cas9系统改造酿酒酵母的研究进展  被引量:2

Advances in CRISPR/Cas9 System Modifying Saccharomyces cerevisiae

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作  者:陈小玲[1] 廖东庆 黄尚飞 陈英[1] 芦志龙[1] 陈东[1] 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)

机构地区:[1]广西科学院非粮生物质能源技术全国重点实验室非粮生物质酶解国家重点实验室,南宁530007 [2]广西轻工业科学技术研究院有限公司,南宁530031

出  处:《生物技术通报》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)。

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.

关 键 词:CRISPR/Cas9系统的构建 多基因编辑 靶序列的设计 脱靶 

分 类 号:Q78[生物学—分子生物学] Q93

 

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