High-efficiency genome editing of an extreme thermophile Thermus thermophilus using endogenous type I and type III CRISPR-Cas systems
作者机构：State Key Laboratory of Agricultural Microbiology and College of Life Science and TechnologyHuazhong Agricultural UniversityWuhanChina Shenzhen Institute of Nutrition and HealthHuazhong Agricultural UniversityShenzhenChina Shenzhen BranchGuangdong Laboratory for Lingnan Modern AgricultureGenome Analysis Laboratory of the Ministry of Agriculture and Rural AffairsAgricultural Genomics Institute at ShenzhenChinese Academy of Agricultural SciencesShenzhenChina College of Biological and Environmental EngineeringXi'an UniversityXi'anChina
出 版 物：《mLife》 (微生物（英文）)
年 卷 期：2022年第1卷第4期
页 面：412-427页
学科分类：0710[理学-生物学] 1007[医学-药学(可授医学、理学学位)] 100705[医学-微生物与生化药学] 07[理学] 071005[理学-微生物学] 10[医学]
基 金：the National Natural Science Foundation of China(32170096) the Fundamental Research Funds for the Central Universities(2662022S KPY001) Cooperation Fund of Huazhong Agricultural University-Agricultural Genomics Institute at Shenzhen(CAAS)(SZYJY2021002)
主 题：endogenous CRISPR-Cas system genome editing reporter gene SOD production Thermus thermophilus
摘 要：Thermus thermophilus is an attractive species in the bioindustry due to its valuable natural products,abundant thermophilic enzymes,and promising fermentation ***,efficient and versatile genome editing tools are not available for this *** this study,we developed an efficient genome editing tool for *** HB27 based on its endogenous type IB,I-C,and III-A/B CRISPR-Cas ***,we systematically characterized the DNA interference capabilities of the different types of the native CRISPR-Cas systems in *** *** found that genomic manipulations such as gene deletion,mutation,and in situ tagging could be easily implemented by a series of genome-editing plasmids carrying an artificial self-targeting mini-CRISPR and a donor DNA responsible for the recombinant *** also compared the genome editing efficiency of different CRISPR-Cas systems and the editing plasmids with donor DNAs of different ***,we developed a reporter gene system for *** based on a heat-stableβ-galactosidase gene TTP0042,and constructed an engineered strain with a high production capacity of superoxide dismutases by genome modification.
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