In Vivo Genome Editing in Type I and II Methanotrophs Using a CRISPR/Cas9 System.

ABSTRACT: Methanotrophic bacteria are Gram-negative, aerobic organisms that use methane as their sole source of carbon and energy. In this study, we constructed and exemplified a CRISPR/Cas9 genome editing system and used it to successfully make gene deletions and insertions in the type I methanotroph Methylococcus capsulatus Bath and the type II methanotroph Methylocystis parvus OBBP. High frequencies of gene deletions and insertions were achieved in combination with homology-directed repair. In M. parvus OBBP, we also investigated the impact of several parameters on the CRISPR/Cas9 genome editing, where the ligD gene was targeted with various PAM sequences and guide RNA spacer sequences, homology arms of variable length, differences in the duration of mating during conj

SUBMITTER: Rumah BL 

PROVIDER: S-EPMC9942187 | biostudies-literature | 2023 Feb

REPOSITORIES: biostudies-literature