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A modular genetic toolbox for precise gene regulation and multi-color imaging in streptococci
Johann Mignolet1, Albane Schmid1, Johan Staub2
1Department of Fundamental Microbiology, Faculty of Biology and Medicine, University of Lausanne, CH-1015 Lausanne, Switzerland.
Microlife
|March 16, 2026
Summary
This study introduces versatile fluorescent protein tools for live imaging in diverse streptococcal species. These new methods enable multicolor labeling, advancing research in bacterial cell biology and infectious disease mechanisms.
Area of Science:
- Microbiology
- Molecular Biology
- Bacterial Genetics
Background:
- Fluorescent labeling is crucial for live cell imaging in microbiology, aiding the study of cellular processes, gene expression, and protein localization.
- While multicolor imaging is common in *Streptococcus pneumoniae* and *S. mutans*, it's less established in other streptococcal species, limiting research scope.
- A need exists for expanded molecular tools to facilitate advanced imaging in a broader range of streptococci.
Purpose of the Study:
- To benchmark fluorescent proteins for use in streptococcal species.
- To develop versatile, transferable fluorescent reporter cassettes for multicolor labeling.
- To enable advanced single-cell imaging strategies in the streptococcus clade.
Main Methods:
- Five fluorescent proteins were fused to the *S. pneumoniae* HlpA protein.
- Reporter cassettes were engineered with antibiotic resistance genes and inducible/constitutive expression systems.
- Methods were developed for transferring these cassettes to *S. salivarius*, *S. thermophilus*, and *S. pyogenes*.
Main Results:
- Successful benchmarking of five fluorescent proteins for streptococcal applications.
- Creation of versatile, transferable fluorescent reporter cassettes.
- Demonstration of triple fluorescent labeling in *S. salivarius* by tagging HlpA, FtsZ, and DivIVA.
Conclusions:
- The developed fluorescent protein tools and cassettes expand the molecular toolbox for streptococcal research.
- These innovations facilitate multicolor single-cell labeling strategies in diverse streptococcal species.
- The availability of these tools will broaden the application of advanced imaging for studying bacterial processes and pathogenesis.
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