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...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2022-12-01
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| Series: | Engineering Microbiology |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2667370322000376 |
| _version_ | 1797788390053117952 |
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| author | Wentao Zheng Yandong Xia Xue Wang Shiqing Gao Diao Zhou Jun Fu Ruijuan Li Jia Yin |
| author_facet | Wentao Zheng Yandong Xia Xue Wang Shiqing Gao Diao Zhou Jun Fu Ruijuan Li Jia Yin |
| author_sort | Wentao Zheng |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-13T01:35:54Z |
| format | Article |
| id | doaj.art-71ac2b3a2e704d9a80876cc1bcbb61ac |
| institution | Directory Open Access Journal |
| issn | 2667-3703 |
| language | English |
| last_indexed | 2024-03-13T01:35:54Z |
| publishDate | 2022-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Engineering Microbiology |
| spelling | doaj.art-71ac2b3a2e704d9a80876cc1bcbb61ac2023-07-04T05:11:24ZengElsevierEngineering Microbiology2667-37032022-12-0124100046Cascade-Cas3 facilitates high-accuracy genome engineering in Pseudomonas using phage-encoded homologous recombinationWentao Zheng0Yandong Xia1Xue Wang2Shiqing Gao3Diao Zhou4Jun Fu5Ruijuan Li6Jia Yin7Shandong University-Helmholtz Institute of Biotechnology, State Key Laboratory of Microbial Technology, Qingdao 266237, ChinaHunan 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; 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, ChinaShandong University-Helmholtz Institute of Biotechnology, State Key Laboratory of Microbial Technology, Qingdao 266237, ChinaShandong University-Helmholtz Institute of Biotechnology, State Key Laboratory of Microbial Technology, Qingdao 266237, ChinaHunan 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, ChinaShandong University-Helmholtz Institute of Biotechnology, State Key Laboratory of Microbial Technology, Qingdao 266237, China; Corresponding authors.Shandong University-Helmholtz Institute of Biotechnology, State Key Laboratory of Microbial Technology, Qingdao 266237, China; Corresponding authors.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; Corresponding authors.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.http://www.sciencedirect.com/science/article/pii/S2667370322000376Phage-encoded homologous recombinationPseudomonasCascade-Cas3 systemGenome engineering |
| spellingShingle | Wentao Zheng Yandong Xia Xue Wang Shiqing Gao Diao Zhou Jun Fu Ruijuan Li Jia Yin Cascade-Cas3 facilitates high-accuracy genome engineering in Pseudomonas using phage-encoded homologous recombination Engineering Microbiology Phage-encoded homologous recombination Pseudomonas Cascade-Cas3 system Genome engineering |
| title | Cascade-Cas3 facilitates high-accuracy genome engineering in Pseudomonas using phage-encoded homologous recombination |
| title_full | Cascade-Cas3 facilitates high-accuracy genome engineering in Pseudomonas using phage-encoded homologous recombination |
| title_fullStr | Cascade-Cas3 facilitates high-accuracy genome engineering in Pseudomonas using phage-encoded homologous recombination |
| title_full_unstemmed | Cascade-Cas3 facilitates high-accuracy genome engineering in Pseudomonas using phage-encoded homologous recombination |
| title_short | Cascade-Cas3 facilitates high-accuracy genome engineering in Pseudomonas using phage-encoded homologous recombination |
| title_sort | cascade cas3 facilitates high accuracy genome engineering in pseudomonas using phage encoded homologous recombination |
| topic | Phage-encoded homologous recombination Pseudomonas Cascade-Cas3 system Genome engineering |
| url | http://www.sciencedirect.com/science/article/pii/S2667370322000376 |
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