Comprehensive double-mutant analysis of the <i>Bacillus subtilis</i> envelope using double-CRISPRi.

ABSTRACT: Understanding bacterial gene function remains a major biological challenge. Double-mutant genetic interaction (GI) analysis addresses this challenge by uncovering the functional partners of targeted genes, allowing us to associate genes of unknown function with novel pathways and unravel connections between well-studied pathways, but is difficult to implement at the genome-scale. Here, we develop and use double-CRISPRi to systematically quantify genetic interactions at scale in the Bacillus subtilis envelope, including essential genes. We discover > 1000 known and novel genetic interactions. Our analysis pipeline and experimental follow-ups reveal the distinct roles of paralogous genes such as the mreB and mbl actin homologs, and identify new genes involved in the well

SUBMITTER: Koo BM 

PROVIDER: S-EPMC11343205 | biostudies-literature | 2024 Aug

REPOSITORIES: biostudies-literature