CRISPR/Cas9-based Genome Editing in Pseudomonas aeruginosa and Cytidine Deaminase-Mediated Base Editing in Pseudomonas Species

• DOI: 10.1016/j.isci.2018.07.024
• Publication Date: 2018-08-01T16:45:43Z
• Authors: Weizhong Chen; Ya Zhang; Yifei Zhang; Yishuang Pi; Tongnian Gu; Liqiang Song; Yu Wang; Quanjiang Ji
• Keywords: Pseudomonas fluorescens; Pseudomonas putida

Pseudomonas species are a large class of gram-negative bacteria that exhibit significant biomedical, ecological, and industrial importance. Despite the extensive research wide applications, genetic manipulation in species, particular major human pathogen aeruginosa, remains laborious endeavor. Here we report development genome editing method pCasPA/pACRISPR by harnessing CRISPR/Cas9 phage λ-Red recombination systems. The allows for efficient scarless P. aeruginosa. By engineering fusion cytidine deaminase APOBEC1 Cas9 nickase, further develop base system pnCasPA-BEC, which enables highly gene inactivation point mutations variety such as putida, fluorescens, syringae. Application two methods will dramatically accelerate investigations, bacterial physiology study, drug target exploration, metabolic engineering.