Cascade-Cas3 facilitates high-accuracy genome engineering in Pseudomona s using phage-encoded homologous recombination  

作　　者：Wentao Zheng Yandong Xia Xue Wang Shiqing Gao Diao Zhou Jun Fu Ruijuan Li Jia Yin 

机构地区：[1]Shandong University-Helmholtz Institute of Biotechnology,State Key Laboratory of Microbial Technology,Qingdao 266237,China [2]Hunan Provincial Key Laboratory of Animal Intestinal Function and Regulation,Hunan International Joint Laboratory of Animal Intestinal Ecology and Health,College of Life Sciences,Hunan Normal University,Changsha 410081,China [3]College of Life Science and Technology,Hunan Provincial Key Laboratory for Control of Forest Diseases and Pests,Key Laboratory for Non-wood Forest Cultivation and Conservation of Ministry of Education,Central South University of Forestry and Technology,Changsha 410004,China

出　　处：《Engineering Microbiology》2022年第4期64-67,共4页工程微生物学（英文）

基　　金：supported by grants from the National Key R&D Program of China(2019YFA0904000);the National Natural Science Foundation of China(31570094 and 81502962);the 111 Project(B16030);the Science and Technology Development Program of Suzhou(SYG201507);the Natural Science Foundation of Jiangsu Province(BK20160368);the Key Programs of Frontier Scientific Research of the Chinese Academy of Sciences(QYZDY-SSW-SMC008);the State Key Laboratory of Microbial Technology Open Projects Fund(M2017-05);the Shandong Provincial Natural Science Foundation of China(ZR2020MC015)to R.L.;the Huxiang Youth Excellent(2017RS3029)to J.Y.;the Taishan Scholar Program of Shandong Province to J.F.

摘　　要：Phage-encoded homologous recombination(PEHR)is an efficient tool for bacterial genome editing.We previously developed and utilized a Pseudomonas-specific PEHR system.However,when using the PEHR system for Pseu-domonas genome editing,false positives can be a problem.In this study,we combined a compact Cascade-Cas3 system from P.aeruginosa(PaeCas3c)with a Pseudomonas-specific PEHR system,and the results of our recom-bineering assay showed that this compact Cascade-Cas3 system can significantly improve PEHR recombineering accuracy.

关 键 词：Phage-encoded homologous recombination PSEUDOMONAS Cascade-Cas3 system Genome engineering 

分 类 号：Q812[生物学—生物工程] Q93