Genetically Encoded RNA-Based Bioluminescence Resonance Energy Transfer (BRET) Sensors

Area of Science:

• Molecular Biology
• Biotechnology
• Nanotechnology

Background:

• RNA nanostructures are increasingly used for biosensors and genetic regulators.
• Current RNA nanodevices often rely on fluorescence, which has limitations like high background and phototoxicity.
• Real-time bioluminescent RNA reporters are underdeveloped.

Purpose of the Study:

• To develop the first genetically encoded RNA-based bioluminescence resonance energy transfer (BRET) sensors.
• To enable real-time target detection in living cells using a bioluminescent readout.
• To provide a modular system for imaging and detecting various cellular analytes.

Main Methods:

• Coupling a luciferase bioluminescence donor with a fluorogenic RNA-based acceptor.
• Engineering modular RNA nanostructures for BRET.
• Testing the system for real-time detection in living cells.

Main Results:

• Demonstrated the first genetically encoded RNA-based BRET sensors.
• Successfully achieved real-time target detection in living cells.
• Showcased the modularity of the BRET system for detecting diverse analytes.

Conclusions:

• The developed RNA-based BRET sensors offer a novel bioluminescent alternative to fluorescence-based systems.
• This technology has broad potential applications in bioanalysis and nanomedicine.
• The system can be used for engineering biosensors, studying RNA-protein interactions, and high-throughput screening or in vivo imaging.