Chen Yibao, Yan Bingjie, Chen Weizhong, Zhang Xue, Liu Zhengjie, Zhang Qing, Li Lulu, Hu Ming, Zhao Xiaonan, Xu Xiaohui, Lv Qianghua, Luo Yanbo, Cai Yumei, Liu Yuqing
Shandong Key Laboratory of Animal Disease Control and Breeding, Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences, Jinan, China.
China-UK Joint Laboratory of Bacteriophage Engineering, Jinan, China.
iScience. 2024 Jun 6;27(7):110210. doi: 10.1016/j.isci.2024.110210. eCollection 2024 Jul 19.
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 phages. We first evaluated the active cutting function of the CRISPR-Cas12a system and discovered that it had a higher gene-cutting efficiency than the type II CRISPR-Cas9 system in three different phages. We also demonstrated the system's ability to precisely edit genes in phages, phages, and phages. Using the aforementioned strategies, non-essential 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.
是一种常见的机会致病菌。噬菌体疗法的潜在疗效已引起研究人员的关注，但缺乏高效的基因编辑工具，限制了对其生物学特性的研究。在此，我们设计了一种用于噬菌体基因编辑的V型CRISPR-Cas12a系统。我们首先评估了CRISPR-Cas12a系统的活性切割功能，发现在三种不同的噬菌体中，它比II型CRISPR-Cas9系统具有更高的基因切割效率。我们还证明了该系统在噬菌体、噬菌体和噬菌体中精确编辑基因的能力。使用上述策略，可以有效删除非必需的噬菌体基因，导致最多减少5215 bp（7.05%）。我们的研究提供了一种快速、高效且省时的工具，加速了噬菌体工程的进展。
