{"title":"Cascade-Cas3利用噬菌体编码的同源重组促进假单胞菌的高精度基因组工程","authors":"Wentao Zheng, Yandong Xia, Xue Wang, Shiqing Gao, Diao Zhou, Jun Fu, Ruijuan Li, Jia Yin","doi":"10.1016/j.engmic.2022.100046","DOIUrl":null,"url":null,"abstract":"<p><p>Phage-encoded homologous recombination (PEHR) is an efficient tool for bacterial genome editing. We previously developed and utilized a <i>Pseudomonas</i>-specific PEHR system. However, when using the PEHR system for <i>Pseudomonas</i> genome editing, false positives can be a problem. In this study, we combined a compact Cascade-Cas3 system from <i>P. aeruginosa</i> (PaeCas3c) with a <i>Pseudomonas</i>-specific PEHR system, and the results of our recombineering assay showed that this compact Cascade-Cas3 system can significantly improve PEHR recombineering accuracy.</p>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"10 1","pages":"100046"},"PeriodicalIF":0.0000,"publicationDate":"2022-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11611027/pdf/","citationCount":"0","resultStr":"{\"title\":\"Cascade-Cas3 facilitates high-accuracy genome engineering in <i>Pseudomona</i>s using phage-encoded homologous recombination.\",\"authors\":\"Wentao Zheng, Yandong Xia, Xue Wang, Shiqing Gao, Diao Zhou, Jun Fu, Ruijuan Li, Jia Yin\",\"doi\":\"10.1016/j.engmic.2022.100046\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Phage-encoded homologous recombination (PEHR) is an efficient tool for bacterial genome editing. We previously developed and utilized a <i>Pseudomonas</i>-specific PEHR system. However, when using the PEHR system for <i>Pseudomonas</i> genome editing, false positives can be a problem. In this study, we combined a compact Cascade-Cas3 system from <i>P. aeruginosa</i> (PaeCas3c) with a <i>Pseudomonas</i>-specific PEHR system, and the results of our recombineering assay showed that this compact Cascade-Cas3 system can significantly improve PEHR recombineering accuracy.</p>\",\"PeriodicalId\":100478,\"journal\":{\"name\":\"Engineering Microbiology\",\"volume\":\"10 1\",\"pages\":\"100046\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2022-08-29\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11611027/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Engineering Microbiology\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1016/j.engmic.2022.100046\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2022/12/1 0:00:00\",\"PubModel\":\"eCollection\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Engineering Microbiology","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1016/j.engmic.2022.100046","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2022/12/1 0:00:00","PubModel":"eCollection","JCR":"","JCRName":"","Score":null,"Total":0}
Cascade-Cas3 facilitates high-accuracy genome engineering in Pseudomonas using phage-encoded homologous recombination.
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 Pseudomonas 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 recombineering assay showed that this compact Cascade-Cas3 system can significantly improve PEHR recombineering accuracy.
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