How we design DNA

Kyle Taylor from the Glowing Plant project explains how we design our DNA sequences

[@LDerby]

You have made cloning immensely easy to understand, bravo

[@germandesigner]

Great idea making the parts of the DNA structure modular!

[@mdickson402]

Glowing science in black and white.... Beautiful

[@MeliponiculturaenCostaRica]

Very precise concepts in a simple way to learn them

[@blazearmoru]

This reminds me of the Gene Compiler I found out about from one of your videos. Please keep us updated! I wana play with that one day ^~^

[@louiswatrin1447]

I absolutely love the work you are doing and I am really passionate about it too. I adore your videos, you explain really well and easily what you are doing. Continue like that👍

[@kyletaylor1810]

Good question. While we will use ligation free cloning for some things, we have settle upon the Golden Braid system. This way we can use one restriction restriction enzyme in a one pot reaction for most/all of our reactions rather than ordering primers for every construct.

[@TAXABiotechnologies]

We can do localized emissions by selecting different promotors, but for now we are focused on making the whole thing glow.

[@VidMasterZ]

I don't wanna see it on a board, I wanna see it at the bench!

[@TAXABiotechnologies]

We don't print all in one go because it means we can swap parts out to try different promotors etc. It's very expensive to print the whole sequence each time!

[@jensstaal]

OK good to know - just that agrobacterium most likely would be easier to set up if DIY labs is the aim... (especially since gene gun methods usually means that you need to regenerate from callus cultures, requiring quite a lot of equipment). Any thoughts about selectable markers to be used?

[@TAXABiotechnologies]

We will be doing both chloroplast and nucleus integration to see which works best

[@gooey342]

have you figured out a way to do localized emissions in the plants, or will the entire thing glow?

[@JasonKnight]

So what technology/process exactly is it that allows you to use one restriction enzyme but have the modules self assemble? And how does this make it so you can swap out modules after the whole vector is built? And why can't you just synthesize the whole vector from scratch from a place like Cambrian Genomics since the sequences of all the components you mentioned have already been sequenced?

[@kyletaylor1810]

We will be using the Golden Braid system. Here are citations to two papers on the system: GoldenBraid: An Iterative Cloning System for Standardized Assembly of Reusable Genetic Modules, PLoS One, July 7, 2011 GoldenBraid 2.0: A Comprehensive DNA Assembly Framework for Plant Synthetic Biology, Plant Physiology, July 13, 2013 vol 162 no 3 If there's interests, we could do a video on the Golden Braid system or do an interview with the lab that developed it. Let us know!

[@juliembs975]

And is the vector a bacterium ??

[@TAXABiotechnologies]

DNA design happens on a computer. Once the DNA has been delivered we will show you the bench work!

[@juliembs975]

Is trna the same as tRNA ? which is "Transfer RNA"

[@StephenHurst1987]

Why not use ligation free cloning instead of dealing with restriction enzyme sites, may be a lot simpler.

[@jensstaal]

Since you are targetting the chloroplast I guess you are stuck with transformation strategies like the gene gun - or are there now modified Agrobacterium Ti plasmids that go to chloroplasts rather than the nucleus? Alternatively - would the lux operon work in the nucleus if each gene was flanked with the self-processing peptide 2A?

[@aljenembtry7781]

Could you take a frilled lizard 🦎🤔make it 3 foot tall and give it spitting cobra faings and glands??? Wouldn't that be a alternative route for DINOSAURS???