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Tunable Expression Tools Enable Single-Cell Strain Distinction in the Gut Microbiome.
Weston R Whitaker1, Elizabeth Stanley Shepherd2, Justin L Sonnenburg2
1Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, CA 94305, USA; Novome Biotechnologies, 100 Kimball Way, South San Francisco, San Francisco, CA 94080, USA.
Cell
|April 22, 2017
Summary
Scientists engineered gut bacteria (Bacteroides) using synthetic biology for improved diagnostics and therapeutics. They developed novel promoters enabling high-level gene expression for imaging and differentiating bacterial species in the gut.
Area of Science:
- Microbiology
- Synthetic Biology
- Gastroenterology
Background:
- Bacteroides are key members of the Western gut microbiota.
- Engineering gut microbes offers potential for diagnostics and therapeutics.
- High-throughput strain modification is crucial for advancing microbiome research.
Purpose of the Study:
- To develop a platform for engineering Bacteroides species.
- To identify novel promoters for high-level gene expression in gut microbes.
- To enable in vivo imaging and differentiation of gut bacteria.
Main Methods:
- Developed a high-throughput strain modification process for Bacteroides.
- Identified and characterized novel phage promoters and translational tuning strategies.
- Engineered fluorescent protein expression for bacterial imaging and species differentiation.
Main Results:
- Achieved unprecedented gene expression levels in engineered Bacteroides.
- Characterized constitutive promoters with a 1,000,000-fold dynamic range and no fitness burden.
- Successfully differentiated six bacterial species in the gut using unique fluorescent signatures.
- Demonstrated that colonization priority dictates colonic crypt occupancy.
Conclusions:
- The developed platform enables robust engineering of Bacteroides for microbiome research.
- Novel promoters provide precise control over gene expression in diverse Bacteroides species.
- Fluorescent tagging allows for real-time monitoring and differentiation of gut microbial populations.
