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Genetic Tagging and Imaging of Proteins with iFAST in Candida albicans
Jonas Devos1, Patrick Van Dijck1, Wouter Van Genechten1
1Laboratory of Molecular Cell Biology, Institute of Botany and Microbiology, KU Leuven, Leuven, Belgium.
Abstract:
Candida albicans is the most common human fungal pathogen, able to reside in a broad range of niches within the human body. Even though C. albicans systemic infection is associated with high mortality, the fungus has historically received relatively little attention, resulting in a lack of optimized molecular and fluorescent tools. Over the last decade, some extra focus has been put on the optimization of fluorescent proteins (FPs) of C. albicans. However, as the FPs are GFP-type, they require an aerobic environment and a relatively long period to fully mature. Recently, we have shown the application of a novel type of fluorogen-based FP, with an improved version of fluorescence activating and absorption shifting tag (iFAST), in C. albicans. Due to the dynamic relation between iFAST and its fluorogens, the system has the advantage of being reversible in terms of fluorescence. Furthermore, the combination of iFAST with different fluorogens results in different spectral and cellular properties, allowing customization of the system. Key features • Genetic integration and tagging with the iFAST tag in Candida albicans. • Imaging and localization of a protein of interest tagged with iFAST. • Reversibility of fluorescence with iFAST.
Insights
This study introduces the novel iFAST system for improved fluorescent protein applications in Candida albicans. This fluorogen-based system offers reversible fluorescence and customizable spectral properties for better fungal research.
Area of Science:
- Microbiology
- Molecular Biology
- Biotechnology
Background:
- Candida albicans is a prevalent human fungal pathogen associated with high mortality rates.
- Existing fluorescent proteins (FPs) for C. albicans, like GFP-type, require aerobic conditions and long maturation times.
- There is a need for optimized molecular and fluorescent tools for C. albicans research.
Purpose of the Study:
- To introduce and evaluate a novel fluorogen-based fluorescent protein system, iFAST, in Candida albicans.
- To demonstrate the advantages of iFAST, including reversibility and customizable spectral properties.
- To enable improved imaging and localization of proteins within C. albicans.
Main Methods:
- Genetic integration and tagging of proteins in Candida albicans using the iFAST tag.
- Utilizing iFAST with various fluorogens to achieve different spectral and cellular properties.
- Investigating the dynamic and reversible nature of iFAST fluorescence.
Main Results:
- Successful genetic integration and tagging of proteins in C. albicans with iFAST.
- Demonstrated imaging and localization capabilities for iFAST-tagged proteins.
- Confirmed the reversible fluorescence characteristic of the iFAST system.
Conclusions:
- The iFAST system provides a versatile and improved tool for fluorescent tagging in Candida albicans.
- Reversible fluorescence and customizable properties of iFAST enhance its utility for fungal research.
- This novel system addresses limitations of traditional FPs in C. albicans studies.
