Advances in CRISPR/Cas9 System Modifying Saccharomycescerevisiae

doi:10.13560/j.cnki.biotech.bull.1985.2022-1534

Abstract

Abstract:

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.

Key words: construction of CRISPR/Cas9 system, multiplex gene editing, design of target sequence, off-target

CHEN Xiao-ling, LIAO Dong-qing, HUANG Shang-fei, CHEN Ying, LU Zhi-long, CHEN Dong. Advances in CRISPR/Cas9 System Modifying Saccharomycescerevisiae[J]. Biotechnology Bulletin, 2023, 39(8): 148-158.

Figures/Tables 3

References 72

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名称 Name	链接 Link	物种Species	优点 Advantage	缺点 Disadvantage	参考文献Reference
GuideScan	-	任何微生物	能以目标酿酒酵母的基因组为参考基因组、有助于减少脱靶位点，设计的靶序列比the mit.edu web interface^[30]、CRISPRscan^[42]及E-CRISP^[56]的好	需在本地计算机安装，需使用命令行，需提供参考基因组	[45]
Yeastriction	http://yeastriction.tnw.tudelft.nl/	酿酒酵母	设计的靶序列比ChopChop^[57]的好，提供33株酿酒酵母的基因组作为备选的参考基因组，不需使用命令行	容易脱靶（除非参考基因组是目标酿酒酵母的基因组）	[26]
CRISPy	http://staff.biosustain.dtu.dk/laeb/crispy_yeast/	酿酒酵母	不需上传参考基因组	只能以酿酒酵母S288C的基因组为参考基因组，为其他酿酒酵母设计靶序列时，容易脱靶	[28]
CRISPy-web	http://crispy.secondarymetabolites.org/	任何微生物	免安装，不需使用命令行，能以目标酿酒酵母的基因组为参考基因组、有助于减少脱靶位点	需上传参考基因组	[55]

名称 Name	链接 Link	物种Species	优点 Advantage	缺点 Disadvantage	参考文献Reference
GuideScan	-	任何微生物	能以目标酿酒酵母的基因组为参考基因组、有助于减少脱靶位点，设计的靶序列比the mit.edu web interface^[30]、CRISPRscan^[42]及E-CRISP^[56]的好	需在本地计算机安装，需使用命令行，需提供参考基因组	[45]
Yeastriction	http://yeastriction.tnw.tudelft.nl/	酿酒酵母	设计的靶序列比ChopChop^[57]的好，提供33株酿酒酵母的基因组作为备选的参考基因组，不需使用命令行	容易脱靶（除非参考基因组是目标酿酒酵母的基因组）	[26]
CRISPy	http://staff.biosustain.dtu.dk/laeb/crispy_yeast/	酿酒酵母	不需上传参考基因组	只能以酿酒酵母S288C的基因组为参考基因组，为其他酿酒酵母设计靶序列时，容易脱靶	[28]
CRISPy-web	http://crispy.secondarymetabolites.org/	任何微生物	免安装，不需使用命令行，能以目标酿酒酵母的基因组为参考基因组、有助于减少脱靶位点	需上传参考基因组	[55]