FluorMango, an RNA-Based Fluorogenic Biosensor for the Direct and Specific Detection of Fluoride.

Area of Science:

• Biotechnology
• Molecular Biology
• Biochemistry

Background:

• Nucleic acids are vital for cellular functions and are attractive for synthetic molecule development.
• Biosensors are crucial for detecting target ligands via measurable signals.
• Fluoride detection is important in various biological and environmental contexts.

Purpose of the Study:

• To introduce FluorMango, a novel fluorogenic ribonucleic acid (RNA)-based biosensor.
• To demonstrate its specificity and high sensitivity for fluoride detection.
• To showcase its application in monitoring defluorinase activity in living bacterial cells.

Main Methods:

• Design of a dual-module RNA biosensor combining a fluoride-specific aptamer (crcB riboswitch) and a light-up RNA (Mango-III).
• Optimization of a communication module using ultrahigh-throughput screening.
• Application of the biosensor for real-time monitoring of defluorinase activity in bacterial cells.

Main Results:

• FluorMango exhibits high fluorescence in the presence of fluoride and low background fluorescence in its absence.
• The biosensor effectively monitors defluorinase activity in living bacterial cells.
• Demonstrated the potential for using FluorMango in future screening for novel defluorinase activities.

Conclusions:

• FluorMango is a highly effective and sensitive RNA-based biosensor for fluoride detection.
• This tool facilitates direct monitoring of defluorinase activity in biological systems.
• FluorMango holds promise for future discovery campaigns in enzyme activity screening.