Polished, long-form articles on synthetic biology topics and my explorations.
The reframed plan acknowledges that endogenous substrate availability is the dominant unknown for firefly bioluminescence in tobacco. Rather than racing to deliver autonomous glow within HTGAA 2026, the plan stages experiments so each milestone produces a real deliverable, with a clear branch point: optimize if the minimum viable construct works, characterize the substrate landscape if it does not.
By searching the firefly proteome for proteins carrying a peroxisome-targeting signal 1 (PTS1), I identified candidates for the two enzymatic steps in the luciferin biosynthesis pathway that remain uncharacterized. This approach bypasses expression-level noise and focuses on subcellular localization as the primary filter.
This is the master reference for my bioluminescence projects: how firefly light works at the molecular level, what we know about luciferin biosynthesis, why no one has achieved autonomous plant glow since Keith Wood's 1986 experiment, and the engineering strategy I'm pursuing to change that.
I used Fallon et al. (2018) lantern expression data to filter the Photinus pyralis genome from 15,773 genes down to a small set of luciferin pathway candidates. After cross-species BLAST, phylogenetics, and manual review, three novel candidates and one literature supported candidate remained.
I found a copper oxidase (PPYR_12315) with ~100× higher expression in the lantern than other copper-binding enzymes. Structure, electrostatics, and conservation all pointed to it as a luciferin synthase candidate. HMMER later showed it was actually a lysyl oxidase aka. an entirely different family. A good lesson in research.