Comprehensive genetic interaction analysis of the Bacillus subtilis envelope using double-CRISPRi.

ABSTRACT: Understanding bacterial gene function remains a major challenge. Double-mutant genetic interaction analysis addresses this challenge by uncovering the functional partners of targeted genes, enabling association of genes of unknown function with known pathways and unraveling of connections among well-studied pathways, but such approaches are difficult to implement at the genome scale. Here, we use double-CRISPR interference (CRISPRi) to systematically quantify genetic interactions at scale for the Bacillus subtilis cell envelope, including essential genes. We discover >1,000 genetic interactions, some known and others novel. Our analysis pipeline and experimental follow-ups reveal the shared and distinct roles of paralogous genes such as mreB and mbl in peptidoglycan and teichoic acid synth

SUBMITTER: Koo BM 

PROVIDER: S-EPMC12716459 | biostudies-literature | 2025 Nov

REPOSITORIES: biostudies-literature