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A Comprehensive, CRISPR-based Functional Analysis of Essential Genes in Bacteria
Open accessAbstract
Summary Essential gene functions underpin the core reactions required for cell viability, but their contributions and relationships are poorly studied in vivo. Using CRISPR interference, we created knockdowns of every essential gene in Bacillus subtilis and probed their phenotypes. Our high-confidence essential gene network, established using chemical genomics, showed extensive interconnections among distantly related processes and identified modes of action for uncharacterized antibiotics. Importantly, mild knockdown of essential gene functions significantly reduced stationary phase survival without affecting maximal growth rate, suggesting that essential protein levels are set to maximize outgrowth from stationary phase. Finally, high-throughput microscopy indicated that cell morphology is relatively insensitive to mild knockdown but profoundly affected by depletion of gene function, revealing intimate connections between cell growth and shape. Our results provide a framework for systematic investigation of essential gene functions in vivo that is broadly applicable to diverse microorganisms and amenable to comparative analysis.
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Cites
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Self-citationGenome-Scale CRISPR-Mediated Control of Gene Repression and Activation
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Self-citationIdentification and characterization of essential genes in the human genome
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Boyd, J. M., Esquilín-Lebrón, K., Campbell, C. J., Kaler, K. R., Norambuena, J., Foley, M. E., Stephens, T. G., Rios, G., Mereddy, G., Zheng, V., et al. (2023) · doi:10.1101/2023.10.03.560778 · Cited by 4
“99 The ylaN gene is essential in Bacillus subtilis and the essentiality can be 100 bypassed by growth with excess Fe salts, but the function of YlaN has not been 101 described (13).”
Background
Metadata
- Authors
- Peters, J. M., Colavin, A., Shi, H., Czarny, T. L., Larson, M. H., Wong, S., Hawkins, J. S., Lu, C. H. S., Koo, B.-M., Marta, E., Shiver, A. L., Whitehead, E. H., Weissman, J. S., Brown, E. D., Qi, L. S., Huang, K. C., & Gross, C. A.
- Venue
- Cell
- Publisher
- Elsevier BV
- Published
- 2016-06
- Volume
- 165
- Issue
- 6
- Pages
- 1493-1506
- ISSN
- 0092-8674 (ISSN-L)
- OpenAlex
- W2407174002
- Cited by
- 714
- References
- 6
- FWCI
- 54.55
- Type
- journal article
- Language
- en
- Open access
- bronze
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