Re: [DIYbio] Increasing production of compounds already present in a plant?

I'm still working, albeit slowly, on my gene guns and will offer more on that as I reach the necessary milestones.

Protoplast transformations and regeneration is more expensive and very skill dependent. Trial and error seems to be the best way to debug protoplast issues. The cell wall degrading enzyme cocktails are expensive. Working on alternative sources as well as a possible recombinant route for said enzymes.

All and all the technical side is very straight forward. The genetic side will, of course, be much trickier. Fishing out whole pathways and expressing that in plants is still a difficult undertaking. A genetic circuit that just makes all the parts all the time does not always lead to increased yield. Fine tuning may be needed. One could start by trying to compare the pathways between orange and coffee/cacao/tea pathways as see where they start to differ and just add the enzymes constitutively to express the missing or different parts. Not sure it will be effective but could also get your hands dirty with plant transforms if time isnt a factor. Primers are fairly cheap (Invitrogen Value Oligos), cloning can get pricey but im sure there are biobricks that can lend a hand. Once you have your circuit, slap on some homologous flanks like the ones used in typical chloroplast transformations like the trnA (alenine) and trnI(isoleucine) intergenic regions and then shoot it into a plant via gene gun or protoplast PEG dna uptake and let the miracle of homologous recombination do all the work. Again, technical side is an endeavor but doable.