Zheng Wentao, Xia Yandong, Wang Xue, Gao Shiqing, Zhou Diao, Fu Jun, Li Ruijuan, Yin Jia
Shandong University-Helmholtz Institute of Biotechnology, State Key Laboratory of Microbial Technology, Qingdao 266237, China.
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.
Eng Microbiol. 2022 Aug 29;2(4):100046. doi: 10.1016/j.engmic.2022.100046. eCollection 2022 Dec.
Phage-encoded homologous recombination (PEHR) is an efficient tool for bacterial genome editing. We previously developed and utilized a -specific PEHR system. However, when using the PEHR system for genome editing, false positives can be a problem. In this study, we combined a compact Cascade-Cas3 system from (PaeCas3c) with a -specific PEHR system, and the results of our recombineering assay showed that this compact Cascade-Cas3 system can significantly improve PEHR recombineering accuracy.
噬菌体编码的同源重组(PEHR)是一种用于细菌基因组编辑的有效工具。我们之前开发并利用了一种特异性的PEHR系统。然而,当使用PEHR系统进行基因组编辑时,假阳性可能会成为一个问题。在本研究中,我们将来自嗜热栖热菌的紧凑型Cascade-Cas3系统(PaeCas3c)与一种特异性的PEHR系统相结合,我们的重组工程分析结果表明,这种紧凑型Cascade-Cas3系统能够显著提高PEHR重组工程的准确性。