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Chemogenetic Modulation of Luciferase Emission Color for Imaging and Sensing
Horgan Manirakiza1, Yuriy Shpinov1, Amandine Gontier1
1Chimie Physique et Chimie du Vivant (CPCV), Sorbonne Université, École Normale Supérieure, Université PSL, CNRS, Paris 75005, France.
ACS Sensors
|April 6, 2026
Summary
Researchers developed LumiFAST, a novel bioluminescent tool for bioimaging. This engineered luciferase offers tunable, red-shifted light emission for enhanced sensitivity and deep tissue imaging, improving cellular visualization.
Area of Science:
- Biotechnology
- Molecular Imaging
- Biochemistry
Background:
- Bioluminescent luciferases are crucial for bioimaging, avoiding external light sources to minimize phototoxicity and background noise.
- Advanced imaging demands luciferases with high photon output, tunable colors, and red-shifted emission for deep tissue penetration.
Purpose of the Study:
- To engineer a small, tunable luciferase (LumiFAST) by fusing NanoLuc with pFAST for advanced bioimaging applications.
- To achieve customizable, red-shifted emission through BRET and explore its utility in live-cell biosensing.
Main Methods:
- Fusion of NanoLuc (blue-light emitter) with pFAST (tunable chemogenetic fluorescent reporter).
- Optimization of fusion topology and linker length to maximize BRET efficiency.
- Characterization of LumiFAST emission spectra and performance in dual-color microscopy and biosensing.
Main Results:
- LumiFAST demonstrates tunable emission from cyan to red by altering fluorogens.
- Achieved high BRET efficiencies up to 90% in the optimized variant.
- Successfully applied LumiFAST for dual-color microscopy and engineered biosensors for protease activity and protein-protein interactions.
Conclusions:
- LumiFAST is a versatile, compact bioluminescent tool enabling tunable, red-shifted emission for enhanced bioimaging.
- The modular design and high BRET efficiency offer significant advantages for multicolor imaging and deep tissue visualization.
- Insights gained facilitate the development of novel NanoLuc-pFAST based biosensors for live-cell analysis.
