[koeng]

I really need to do a write-up. I kinda just whip up the easiest path and do it.

For example for the grape, I needed to knock out some tryptophan synthesis genes so I could redirect the bioflux. Problem is that in bakers yeast they have a whole buncha copies of their chromosomes, so I had to knock out one of the genes and replace it with a different gene from grapes. Did that with a quick lil CRISPR switch.

Had to electroporate tho because the transformation rates on wild/bakers/non-lab yeast are so garbage

[amarant]

How much expensive lab equipment do you need to do something like this?

"A quick lil CRISPR switch" sounds like "oh just my homemade fusion reactor hooked up to my kitchen warp drive" to me, yet you make it sound so simple!

[adastra22]

You can do this on your desk, or better yet at your local diy bio hackerspace. Validating it takes some equipment, e.g. for PCR, but that's commonly available.

[koeng]

Eh, other than the electroporator I could probably do it for about $100-$200 bucks of equipment if I had a decent kitchen.

Reagents probably about $300, but you can use em in a bunch of reactions, in aggregate down to like $50.

The fundamentals of biology are really cheap, but the skills to actually do it are really expensive. It’s way more manual than you imagine - like how my thumb moves. The equipment is way more fundamentally basic than you imagine: the only thing you can’t 3d print and build from off-the-shelf stuff is the instant pot I use for media prep

[Hammerhead12321]

Starting from nothing, how much time (and money) might it take an individual to learn how to be able to do these things?

[koeng]

I’d recommend buying an Odin kit and just trying it. Doesn’t take THAT much to get into genetic engineering.

The tough part is mostly the finesse in the simple things, like trying this in bakers yeast rather than lab yeast, or the genetic design.

Cost is quite high for mistakes, but LLMs are honestly quite good to help you out with the basics. You MUST at least try to read the papers though - it’s not like coding where you can mostly let it do its thing.

[fc417fc802]

If you can find a nearby community college offering a molecular biology class that includes a practical lab then I'd say a couple quarters of time and tuition.

DIY that will depend on your level of ability. You can do this stuff in your kitchen but learning it from a textbook will be daunting for many (most?) people.

[dnautics]

Curious why you do CRISPR when HR works great in yeast.

If you are having trouble transforming, try spheroplasty.

[koeng]

Already answered there: I’m using bakers yeast, not lab yeast (store-bought S cerevisiae). It’s not haploid, often it’s tetraploid. HR doesn’t guarantee homozygous transformation.

Same answer for electroporation vs spheroplasty. I’ve found with wild yeasts or less tamed yeasts (pichia), sometimes just nuking the damn thing with kV will just work, whereas those chemical methods can be way more finicky. Time is money