Prp8 Intein-Targeted Fluorescent Probe for Specific Detection of Cryptococcus neoformans and Inhibitor Screening

Area of Science:

• Medical Mycology
• Biochemistry
• Chemical Biology

Background:

• Cryptococcus neoformans (C. neoformans) infections are a major global health concern, particularly for immunocompromised individuals.
• Developing rapid, cost-effective diagnostics for C. neoformans is challenging, especially in resource-limited settings.
• The Prp8 intein in C. neoformans is a promising target for selective diagnostic probes and antifungal therapies.

Purpose of the Study:

• To develop novel fluorescent probes for the selective detection of C. neoformans.
• To evaluate the efficacy of these probes in vitro and in vivo.
• To establish a rapid screening method for Prp8 intein inhibitors.

Main Methods:

• Synthesis of environment-sensitive fluorescent probes incorporating Prp8 intein inhibitors.
• Evaluation of probe selectivity against Candida species.
• Dual-staining assays combining capsule staining and intracellular Prp8 intein labeling.
• In vivo imaging of fungal infections in a Galleria mellonella model.
• Assessing the dose-dependent relationship between fluorescence intensity and inhibitor activity.

Main Results:

• Probe H2 demonstrated specific detection of C. neoformans with minimal cross-reactivity with Candida species.
• Dual-staining significantly enhanced diagnostic accuracy.
• Successful fluorescence imaging of C. neoformans infections in vivo using the Galleria mellonella model.
• A rapid (2-hour) screening assay for Prp8 intein inhibitors was established based on fluorescence intensity.

Conclusions:

• Fluorescent probe H2 offers a selective and sensitive method for C. neoformans detection.
• The developed probes and dual-staining approach can improve the diagnosis of cryptococcosis.
• The probe facilitates efficient in vivo imaging and rapid screening of novel antifungal agents targeting the Prp8 intein.