Brominated Luciferins Are Versatile Bioluminescent Probes

Area of Science:

• Chemical synthesis
• Biotechnology
• Molecular imaging

Background:

• Luciferins are essential substrates for bioluminescence imaging.
• Development of novel luciferin analogues can enhance imaging sensitivity and specificity.
• Brominated luciferins offer unique photophysical properties for investigation.

Purpose of the Study:

• To synthesize and characterize a series of brominated luciferin analogues.
• To evaluate the performance of these analogues in bioluminescence imaging applications.
• To explore their potential as substrates for firefly luciferase and orthogonal mutant enzymes.

Main Methods:

• Common synthetic route for regioisomeric brominated luciferin scaffolds.
• Bioluminescence imaging assays using firefly luciferase.
• Computational analysis and photophysical measurements of photon output.
• In vitro cell-based assays and in vivo animal model evaluations.

Main Results:

• Successful synthesis of diverse brominated luciferin analogues.
• Varying light emission levels observed across analogues with firefly luciferase.
• The brightest analogue demonstrated enhanced performance over native luciferin in cellular models at low doses.
• Other analogues, while weak emitters with firefly luciferase, showed potential for orthogonal mutant enzymes.

Conclusions:

• Brominated luciferins represent a promising class of compounds for bioluminescence imaging.
• Specific analogues can offer improved sensitivity and performance in biological systems.
• The developed analogues hold potential for applications with both native and engineered luciferase systems.