ArticleCell systems2025
Comprehensive genetic interaction analysis of the Bacillus subtilis envelope using double-CRISPRi.
Article in Cell systems, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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14 citing papers in PubMed.
- The phenotypic landscape of the model firmicuteProceedings of the National Academy of Sciences of the United States of America · 2026Article
- Elevated temperature undermines the efficacy ofmSphere · 2026Article
- Applications of transposon-insertion sequencing for understanding bacterial physiology.Microbial genomics · 2026Review
- A broadly conserved gram-positive lipoprotein regulates cell elongation.Proceedings of the National Academy of Sciences of the United States of America · 2026Article
- The genetic landscape of antibiotic sensitivity inScience advances · 2026Article
- Systematic CRISPRi screening reveals genetic modulators ofbioRxiv : the preprint server for biology · 2026Article
- Post-translational modifications of GlmR integrate metabolic and stress signals to maintain cell envelope homeostasis in Bacillus subtilis.PLoS genetics · 2026Article
- Elucidating the roles of essential genes in autotrophic metabolism and cell morphology of Clostridium ljungdahlii by CRISPRi.Applied microbiology and biotechnology · 2026Article
- Multi-omics profiling reveals atypical sugar utilization and a key membrane composition regulator in Streptococcus pneumoniae.Nature communications · 2025Article
- Staying in the loop to make ends meet: roles and regulation of GlmR inbioRxiv : the preprint server for biology · 2025Article
- RagB stimulates the activity of the peptidoglycan polymerase RodA in Bacillus subtilis.EMBO reports · 2025Article
- Considering Metabolic Context in Enzyme Evolution and Design.Biochemistry · 2025Review
- Peering into the Bacterial Cell: From Transcription to Functional Genomics.Journal of molecular biology · 2025Review
- Correction of a widespread bias in pooled chemical genomics screens improves their interpretability.Molecular systems biology · 2024Article
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Authors and funding
10 authors.
Funding
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 synthesis and identify additional genes involved in the well-studied process of cell division. Overall, our study provides valuable insights into gene function and demonstrates the utility of double-CRISPRi for high-throughput dissection of bacterial gene networks, providing a blueprint for future studies in diverse species. A record of this paper's transparent peer review process is included in the supplemental information.
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