Development of the CRISPR-Cas12a system for editing of Pseudomonas aeruginosa phages

文献类型： 外文期刊

第一作者： Chen, Yibao

作者： Chen, Yibao;Yan, Bingjie;Zhang, Xue;Liu, Zhengjie;Zhang, Qing;Li, Lulu;Hu, Ming;Zhao, Xiaonan;Xu, Xiaohui;Lv, Qianghua;Luo, Yanbo;Liu, Yuqing;Chen, Yibao;Yan, Bingjie;Zhang, Xue;Liu, Zhengjie;Zhang, Qing;Li, Lulu;Hu, Ming;Zhao, Xiaonan;Xu, Xiaohui;Lv, Qianghua;Luo, Yanbo;Liu, Yuqing;Chen, Weizhong;Cai, Yumei;Chen, Yibao;Hu, Ming;Liu, Yuqing

作者机构：

期刊名称：ISCIENCE （ 影响因子：4.6； 五年影响因子：5.0 ）

ISSN：

年卷期： 2024 年 27 卷 7 期

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收录情况： SCI

摘要： Pseudomonas aeruginosa is a common opportunistic pathogen. The potential efficacy of phage therapy has attracted the attention of researchers, but efficient gene-editing tools are lacking, limiting the study of their biological properties. Here, we designed a type V CRISPR-Cas12a system for the gene editing of P. aeruginosa phages. We first evaluated the active cutting function of the CRISPR-Cas12a system in vitro and discovered that it had a higher gene-cutting efficiency than the type II CRISPR-Cas9 system in three different P. aeruginosa phages. We also demonstrated the system's ability to precisely edit genes in Escherichia coli phages, Salmonella phages, and P. aeruginosa phages. Using the aforementioned strategies, non-essential P. aeruginosa phage genes can be efficiently deleted, resulting in a reduction of up to 5,215 bp (7.05%). Our study has provided a rapid, efficient, and time-saving tool that accelerates progress in phage engineering.

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