Developing a CRISPR-assisted base-editing system for genome engineering of Pseudomonas chlororaphis.

ABSTRACT: Pseudomonas chlororaphis is a non-pathogenic, plant growth-promoting rhizobacterium that secretes phenazine compounds with broad-spectrum antibiotic activity. Currently available genome-editing methods for P. chlororaphis are based on homologous recombination (HR)-dependent allelic exchange, which requires both exogenous DNA repair proteins (e.g. λ-Red-like systems) and endogenous functions (e.g. RecA) for HR and/or providing donor DNA templates. In general, these procedures are time-consuming, laborious and inefficient. Here, we established a CRISPR-assisted base-editing (CBE) system based on the fusion of a rat cytidine deaminase (rAPOBEC1), enhanced-specificity Cas9 nickase (eSpCas9pp^D10A ) and uracil DNA glycosylase inhibitor (UGI). This CBE system converts C:G into T:A with

SUBMITTER: Yue SJ 

PROVIDER: S-EPMC9437888 | biostudies-literature | 2022 Sep

REPOSITORIES: biostudies-literature