School of Physical Science and Technology, ShanghaiTech University, Shanghai, China.
Methods Mol Biol. 2022;2479:105-117. doi: 10.1007/978-1-0716-2233-9_8.
CRISPR/Cas9 systems have been widely adopted for genetic manipulation in diverse biological systems owing to the ease of use and high efficiency. We have recently developed a CRISPR/Cas9-based genome editing system (pCasKP-pSGKP) by coupling a CRISPR/Cas9 system with the lambda Red recombination system as well as a cytidine deaminase-mediated base editing system (pBECKP) in Klebsiella pneumoniae, enabling rapid, scarless, and efficient genetic manipulation in diverse K. pneumoniae strains. In this chapter, we introduce the detailed procedures of using these two tools for genome editing in K. pneumoniae.
CRISPR/Cas9 系统由于其使用简便和高效，已经在各种生物系统中被广泛用于遗传操作。我们最近通过将 CRISPR/Cas9 系统与 lambda Red 重组系统以及胞嘧啶脱氨酶介导的碱基编辑系统（pBECKP）相结合，在肺炎克雷伯氏菌中开发了一种基于 CRISPR/Cas9 的基因组编辑系统（pCasKP-pSGKP），从而能够在各种肺炎克雷伯氏菌菌株中实现快速、无痕和高效的遗传操作。在本章中，我们将介绍使用这两种工具在肺炎克雷伯氏菌中进行基因组编辑的详细步骤。
