Authors
Lili M Kim, Byoung-Mo Koo, Hannah N Burkhart, Daniela Vollmer, Waldemar Vollmer, Horia Todor, Carol A Gross
Published in
Proceedings of the National Academy of Sciences of the United States of America. Volume 123. Issue 40. Pages e2619440123. Oct 06, 2026. Epub Sep 29, 2026.
Abstract
Firmicutes are gram-positive bacteria with important roles in human health, disease, and industry. However, more than a quarter of genes in the model firmicute Bacillus subtilis remain completely uncharacterized, including numerous phylum-specific genes. Here, we design a compact pooled CRISPRi library targeting all protein-coding genes in B. subtilis and test its growth in ~150 stress conditions. Using data from this screen as a hypothesis generator, we perform targeted experiments, revealing that the conserved essential firmicute protein YneF is part of the SRP cotranslational protein secretion pathway. We also demonstrate that ECF-transporters play a broadly conserved role in cell wall homeostasis, perform an unbiased analysis of amino acid crossfeeding, and make additional insights into bacterial competition. This work additionally provides a rich dataset to nucleate future studies of uncharacterized genes and presents a framework for performing accessible full-genome functional genomic screens in other bacteria.
PMID:
42809408
Bibliographic data and abstract were imported from PubMed on 30 Sep 2026.
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