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...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.展开更多
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)+7 种基金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.
文摘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.
