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In vivo Dual Substrate Bioluminescent Imaging
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BODIPY-coelenterazine conjugates as self-illuminating substrates for NanoLuc
Valeska Viereckt1, Frank Abendroth1, Alexander Schauerte2
1Department of Chemistry, Marburg University, Marburg, Germany. olalla.vazquez@staff.uni-marburg.de.
Chemical Communications (Cambridge, England)
|March 12, 2025
Summary
We conjugated coelenterazine (CTZ) to BODIPY, creating a molecule that retains enzyme activity and offers fluorescent properties. This advance enables new applications like bioluminescence imaging-guided photodynamic therapy.
Area of Science:
- Chemical Biology
- Biochemistry
- Photochemistry
Background:
- Luciferase enzymes catalyze light-producing reactions using substrates like coelenterazine (CTZ).
- BODIPY dyes are known for their fluorescence and ability to generate singlet oxygen (¹O₂).
- Combining these functionalities could lead to novel theranostic agents.
Purpose of the Study:
- To synthesize and characterize BODIPY-CTZ conjugates.
- To evaluate the enzymatic activity of BODIPY-CTZ with luciferase.
- To explore the potential of these conjugates in applications requiring bioluminescence and singlet oxygen generation.
Main Methods:
- Chemical synthesis of coelenterazine conjugated to BODIPY.
- Enzymatic assays using luciferase-type enzymes to assess substrate activity.
- Spectroscopic analysis to confirm BODIPY's fluorescent properties and singlet oxygen generation capacity.
Main Results:
- Successful conjugation of coelenterazine to BODIPY was achieved.
- The resulting BODIPY-CTZ conjugate maintained its activity as a substrate for luciferase enzymes.
- The conjugate exhibited retained BODIPY fluorescence and singlet oxygen generation capabilities.
Conclusions:
- BODIPY-CTZ conjugates represent a novel class of molecules with dual functionality.
- These conjugates can serve as versatile substrates for bioluminescent reporters and photosensitizers.
- Potential applications include advanced imaging techniques and targeted therapies like photodynamic therapy.
