Genome editing and transcriptional repression in Pseudomonas putida KT2440 via the type II CRISPR system.

ABSTRACT: BACKGROUND:The soil bacterium Pseudomonas putida KT2440 is a "generally recognized as safe"-certified strain with robust property and versatile metabolism. Thus, it is an ideal candidate for synthetic biology, biodegradation, and other biotechnology applications. The known genome editing approaches of Pseudomonas are suboptimal; thus, it is necessary to develop a high efficiency genome editing tool. RESULTS:In this study, we established a fast and convenient CRISPR-Cas9 method in P. putida KT2440. Gene deletion, gene insertion and gene replacement could be achieved within 5 days, and the mutation efficiency reached > 70%. Single nucleotide replacement could be realized, overcoming the limitations of protospacer adjacent motif sequences. We also applied nuclease-deficient Cas9 binding at th

SUBMITTER: Sun J 

PROVIDER: S-EPMC5851096 | biostudies-literature | 2018 Mar

REPOSITORIES: biostudies-literature