建立用于铜绿假单胞菌噬菌体高效基因组编辑的CRISPR-Cas12a的方案。
Protocol for establishing CRISPR-Cas12a for efficient genome editing of Pseudomonas aeruginosa phages.
作者信息
Yan Bingjie, Liu Yujia, Cai Yumei, Liu Yuqing, Chen Yibao
机构信息
Shandong Key Laboratory of Animal Disease Control and Breeding, Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences, Jinan, China; College of Veterinary Medicine, Shandong Agricultural University, Taian, China.
College of Veterinary Medicine, Shandong Agricultural University, Taian, China.
出版信息
STAR Protoc. 2024 Dec 20;5(4):103488. doi: 10.1016/j.xpro.2024.103488. Epub 2024 Dec 11.
We developed an efficient type V CRISPR-Cas12a system tailored specifically for Pseudomonas aeruginosa phages, showcasing its remarkable cleavage activity and the ability to precisely introduce genetic modifications, including point mutations, deletions, and insertions, into phage genomes. Here, we present a protocol for establishing CRISPR-Cas12a for genome editing of Pseudomonas aeruginosa phages. We describe steps for the construction of pCRISPR-12a plasmid and guide RNA and the utilization of the type V CRISPR-Cas12a system for precise genetic editing of phages. For complete details on the use and execution of this protocol, please refer to Chen et al..
我们开发了一种专门针对铜绿假单胞菌噬菌体定制的高效V型CRISPR-Cas12a系统，展示了其卓越的切割活性以及将基因修饰（包括点突变、缺失和插入）精确引入噬菌体基因组的能力。在此，我们提供了一种用于建立CRISPR-Cas12a以对铜绿假单胞菌噬菌体进行基因组编辑的方案。我们描述了构建pCRISPR-12a质粒和引导RNA的步骤，以及利用V型CRISPR-Cas12a系统对噬菌体进行精确基因编辑的过程。有关此方案的使用和执行的完整详细信息，请参考Chen等人的研究。
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