DIY genetic engineering - a garage project, how possible is it?

[tantric]

Chill, it's still impossible. I remember hearing in a lecture circa 95 the details of how DNA is organized in fertilized fish eggs and thinking - you can just inject the DNA? No vector no problem? Why don't I have glow in the dark goldfish? Well, it took another decade, but it did happen. In addition to glofish, there are now pink angelfish (angelfish are cichlids, a clade of relatively intelligent fish given to paternal care, much better than danios). I read that it takes 10,000 tries to get a correct insertion, in the angelfish case, but that's also doable....

A bit on CRISPR technology

 

[Malintent]

The person that invents the tools / process to do homebrew DNA manipulation will be the last living billionaire... because it will end the world in a biological Armageddon.

 

[tantric]

okay - fish zygotes have non-nuclear DNA. with a nice micro-needle, you can just inject your new DNA into the egg. you can't control where and if it binds, but it can work. this is basically how the pink angelfish were made. it takes about 10,000 tries to get one that works. how the hell are pink angelfish part of the armageddon?



the current technology involves injecting dyes into fish - see these...

 

[Malintent]

I did an amazon search for "micro-needle' and couldn't find anything relating to injecting DNA.
I also couldn't find anything on Amazon.com for extracting unfertilized eggs from any kind of creature.
additionally, I could not find the "pink skin" DNA kit that I would be injecting into the egg I can't extract, using the micro needle I can't find.

Until such time as it is trivially easy to homebrew DNA experiments, I think we are safe from Armageddon.

the topic was yours. You might be more interested in pink skinned fish than someone else that is more interested in poison secreting gut bacteria, or meth crying alligators... or feminized dinosaurs..

 

[tantric]

as you wish:

Production of transgenic tilapia (Oreochromis niloticus) by one-cell-stage microinjection

Novel genes have been successfully introduced into fish of many species, chiefly by microneedle injection into fertilized eggs. Tilapia are important warm water food fishes with short generation times, but previous attempts to introduce transgenes into this species succeeded only with multicell embryo injection, a procedure that is likely to result in subsequent transgene mosaicism and probable failure of germline transmission to an F1 generation. We here report the successful production of transgenic tilapia (Oreochromis niloticus) by injection of transgenes (mouse metallothionein promoter/rat growth hormone gene) into the one-celled eggs. Slot blotting reveals the presence of transgene copies in many fish, and Southern blotting patterns suggest genomic integration in roughly 20% of fish developing from injected eggs. Most but not all of the transgenic fish appear to be mosaic in that copies are present only in some tissues. In some male fish analysed, this includes sperm. Introduction of DNA constructs in which a fish gene promoter is spliced to a bacterial CAT (chloramphenicol acetyl transferase) gene, together with subsequent CAT assays, were used to test for gene expression in the transgenic fish. A proportion (approximately 30%) gave positive results in these assays.

search Google Scholar for "transgenic fish microinjection"

extracting unfertilized eggs from fish, which normally use external fertilization, is trivial - its what they do. getting the sperm and keeping it alive is more difficult, but there are established protocols. this is how GloFish came to be - danios are lab animals.

Taiwanese engineer first florescent pink angelfish

Green Fluorescent Protein (from jellyfish, there are several colors)

 

[Malintent]

So what is involved in setting up my own 'novel-DNA' production factory in my home?

I am a skilled homebrewer of beer. I grow my own hops, roast my own barley, and I even have a very small yeast bank in the back corner of my refrigerator that I harvested and maintained myself. I built my own recirculating infusion mash system, run by a PID driven SSR that manages infusion temperature to within 0.1 degrees. I operate about 4 times a year and generate less than 50 gallons of beer per year, which I keg, and bottle the same way commercial breweries do.. via counter pressure filling that I built by hand as well.

I'm pretty good at it. Been doing this for a long time. Very easy to get into, but both an art and a science to master.

And all of this is done, literally, in the corner of my detached garage.

So, what do I need to do to expand my setup so that my yeast can be genetically manipulated to have a much higher attenuation (tolerance for alcohol before they drown in their own piss), and a slightly higher flocculation (tendency to sink to the bottom of the fermenter)?

What equipment need I have? Is the fact that my garage is far from completely sanitary an issue? How do I get started?

If you have easy answers for this, then this might possibly be a candidate for 'garage homebrewing'. so far, it does not.

If you want to put this down and pick up what possibly will be the best beer you ever tasted, then read this:

John Palmer's free book, How to Brew Beer (or, as everyone else calls it, the bible)

http://howtobrew.com/book/introduction

Then buy one of these to get you started:

http://www.northernbrewer.com/brewing/beer-equipment-starter-kits

Once you fall in love with the hobby, add more equipment to your system until you end up with something like this:

http://www.theelectricbrewery.com/

THAT's a garage homebrewery

 

[tantric]

okay, no. in the established technology of gengineering there are two avenues of approach that might allow DIY, as i see it. the first is microinjection of fish and amphibian zygotes. fish and friends, when they are just fertilized eggs, have non-nuclear DNA. meaning it's not all packed into chromosomes in the nucleus, it's just floating around in the cytoplasm.the tech, which is proven and established, is to get these zygotes at just the right time and inject bits of DNA into the cell. you can buy the tools for this and the DNA sequences. it's tricky - no joke, it's a specialized skill, but it can be done and has been done. it's just exploiting the preexisting power of nature. oh, and expect 1 out of 10,000 to express your trait and be healthy.

now CRISPR...this is another way in. but it's MUCH more powerful.

your avenue would be to acquire some gengineered yeast strains, like this one, get several cultures, then selectively breed them. this is the state of the art in transgenic yeast

allow me to retract: Biohackers Are Engineering Yeast to Make THC

 

[Loren Pechtel]

The person that invents the tools / process to do homebrew DNA manipulation will be the last living billionaire... because it will end the world in a biological Armageddon.

http://www.johnringo.net/TheLibrary/BlackTideRising/UnderaGraveyardSky.aspx

 

[tantric]

The person that invents the tools / process to do homebrew DNA manipulation will be the last living billionaire... because it will end the world in a biological Armageddon.

http://www.johnringo.net/TheLibrary/BlackTideRising/UnderaGraveyardSky.aspx

okay, chicken little. there are no zombie plagues. creating any kind of recombinant virus is seriously difficult and you CAN'T just order the shit off the net. it would be much easier to make some kind of chemical weapon.

what bugs me is the accepted idea that genetic technology is safe in the hands of industrial agriculture and corporate capitalism. are y'all INSANE? the major advance has been creating crops immune to a certain pesticide so it can be sprayed over the fields by the SHITLOAD. which saves money, apparently. this is NOT responsible use of the technology. industrial agriculture will never use GMO for what it should be used for because they don't make things for poor starving people....but i might.

 

[Malintent]

http://www.johnringo.net/TheLibrary/BlackTideRising/UnderaGraveyardSky.aspx

okay, chicken little. there are no zombie plagues. creating any kind of recombinant virus is seriously difficult and you CAN'T just order the shit off the net. it would be much easier to make some kind of chemical weapon.

what bugs me is the accepted idea that genetic technology is safe in the hands of industrial agriculture and corporate capitalism. are y'all INSANE? the major advance has been creating crops immune to a certain pesticide so it can be sprayed over the fields by the SHITLOAD. which saves money, apparently. this is NOT responsible use of the technology. industrial agriculture will never use GMO for what it should be used for because they don't make things for poor starving people....but i might.

I agree... very dangerous stuff... in the hands of the group, AND in the hands of the individual... at least in the hands of the group, there are some oversights.. some... maybe...

in the hands of an individual.. calamity, terror, and apocalypse.