Multicolor Bioluminescence Obtained Using Firefly Luciferin.

Area of Science:

• Biochemistry
• Molecular Biology
• Biotechnology

Background:

• Firefly bioluminescence is a crucial tool in life science, used for microbial monitoring, cell viability assays, and reporter-gene studies.
• Recent advancements utilize bioluminescence for noninvasive, real-time in vivo imaging, but limitations exist regarding commercial luciferase gene availability and the need for near-infrared (NIR) emission for deep tissue penetration.

Purpose of the Study:

• To review recent advancements in synthetic luciferin analogues for red-shifted bioluminescence.
• To present novel luciferase substrates emitting red, green, and blue light, with a focus on NIR emission for in vivo applications.

Main Methods:

• Literature review of synthetic luciferin analogues with predicted red-shifted bioluminescence.
• Design and synthesis of novel luciferase substrates.
• Characterization of emission wavelengths, including assessment for NIR capabilities.

Main Results:

• Several synthetic luciferin analogues were designed and developed, exhibiting red-shifted bioluminescence.
• The longest emission wavelength achieved was 675 nm, falling within the near-infrared spectrum.
• One synthesized compound is now commercially available as "Aka Lumine®".

Conclusions:

• Synthetic luciferin analogues offer expanded spectral properties for bioluminescence applications.
• The development of NIR-emitting substrates significantly enhances the potential of bioluminescence for in vivo imaging.
• The commercial availability of these novel compounds facilitates their adoption in research settings.