Mobile CRISPR-Cas9 based anti-phage system in E. coli  

作　　者：Zhou Cao Yuxin Ma Bin Jia Ying-Jin Yuan 

机构地区：[1]Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering(Ministry of Education),Tianjin University,Tianjin 300072,China [2]Collaborative Innovation Center of Chemical Science and Engineering(Tianjin),School of Chemical Engineering and Technology,Tianjin University,Tianjin 300072,China

出　　处：《Frontiers of Chemical Science and Engineering》2022年第8期1281-1289,共9页化学科学与工程前沿（英文版）

基　　金：support from the National Key Research and Development Program of China(Grant No.2019YFA0903800);the National Natural Science Foundation of China(Grant Nos.31800719,21621004);the International(regional)cooperation and exchange projects(Grant No.31861143017).

摘　　要：Escherichia coli is one of the most important microbial cell factories,but infection by bacteriophages in the environment may have a huge impact on its application in industrial production.Here,we developed a mobile CRISPR-Cas9 based anti-phage system for bacteriophages defense in E.coli.Two conjugative plasmids pGM1(phosphoglucomutase 1)and pGM2 carrying one and two guide RNAs,respectively,were designed to defend against a filamentous phage.The results showed that the pGM1 and pGM2 could decrease the phage infection rate to 1.6%and 0.2%respectively in infected cells.For preventing phage infection in E.coli,the pGM2 decreased the phage infection rate to 0.1%,while pGM1 failed to block phage infection.Sequence verification revealed that point mutations in protospacer or protospacer adjacent motif sequences of the phage genome caused loss of the defense function.These results support the potential application of MCBAS in E.coli cell factories to defend against phage infections.

关 键 词：phage infections anti-phage CRISPR-Cas9 conjugative transfer synthetic biology 

分 类 号：R378.21[医药卫生—病原生物学]