Coelenterazine and vargulin are naturally-occurring imidazopyrazinone luciferins used in bioluminescence imaging, but their scarcity and need to supply them exogenously to cell culture and animal models limit their use. I propose to elucidate the biosynthesis of these compounds in bioluminescent zooplankton.
In Specific Aim 1, I discuss the possible biosynthetic origins for these molecules, and my plans to identify the genes coding for the requisite biosynthetic enzymes.
In Specific Aim 2, I describe how I would apply these newly-discovered genes to economically produce coelenterazine by fermentation in S. cerevisiae, addressing a critical barrier to progress.
In Specific Aim 3, I propose initial experiments to assess whether the ability to effect the luciferin?s biosynthesis in human cell culture model improves bioluminescence imaging contrast. Successful completion of this project could have revolutionary impacts on biological imaging, akin to how development of Green Fluorescent Protein from a biological curiosity into a multipurpose biochemical marker has radically transformed the way we apply microscopy and imaging to biological questions. Besides improving imaging signal-to-noise, localized in situ luciferin production in cell culture could aid research into inter- and intracellular signaling. Moreover, expression of the luciferin biosynthetic machinery in animals would allow for long-term noninvasive in vivo bioluminescence experiments such as gene expression monitoring and assessing tumor burden in cancer disease models.

Public Health Relevance

Many marine animals glow in the dark, a phenomenon called bioluminescence, but the cellular machinery that produces the molecule responsible for bioluminescence (?luciferin?) is unknown. I propose to identify the genes required for luciferin production in zooplankton, and express these genes in yeast and in human cell culture. If successful, this research has the potential to revolutionize biological imaging in biomedical research, similar to this field?s previous transformation spurred by the biotechnological development of Green Fluorescent Protein.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Postdoctoral Individual National Research Service Award (F32)
Project #
5F32GM129960-03
Application #
9979650
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Hoodbhoy, Tanya
Project Start
2018-08-15
Project End
2021-08-14
Budget Start
2020-08-15
Budget End
2021-08-14
Support Year
3
Fiscal Year
2020
Total Cost
Indirect Cost
Name
University of California, San Diego
Department
Zoology
Type
Earth Sciences/Resources
DUNS #
804355790
City
La Jolla
State
CA
Country
United States
Zip Code
92093