Now, about that gold gun...

[Sebastian S. Cocioba]

How hard would it be to DIY a compressor attached to a thin tube on a pedestal, then shoot tungsten powder downward at some target tissue?

Does anyone have access to any gold gene gun like the PDS-1000 and the fortune of a laxed PI that wont mind some tinkering to test novel delivery methods?

Anyone seriously thinking of attempting a DIY biolisitc device? Would there be massive legal implications if one were to democratize direct genetic engineering much like manufacturing through personal 3d printers?

I know there was an old thread aboot this but I am just being curious since it doesn't seem to be soo crazy of a hack weekend project. Helium is cheap. Using fine enough sand paper would yield very small shot powder from a cube of tungsten that can e bought at McMaster...i think. Or maybe that salt alternative like one DIYBiologist said in the thread.

Sebastian S Cocioba
CEO & Founder
New York Botanics, LLC

Sent via Mobile E-Mail

[Mega]

You can also get tungsten from "ancient" lightbulbs. (meaning not the energy-saving lamps)

The thread which is heated is made of tungsten, although it's not much material at all.


A compressor sucks in air, well, also the contaminants are shot onto the plates. Ok, you select with kanamycin afterwards, which kills *most of* the life formas subjected to it.

[Forrest Flanagan]

It's not terribly hard to manufacture a diaphragm valve, and that would provide a very quick, intense, burst of gas. There's a lot of DIY data about them because they're used in spud guns.

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[Ben Hunt]

The gene gun has its roots in DIY modification of off the shelf materials, (modified crosman air pistol -> nail gun cartridge on a piece of plastic, see wikipedia) but it is a bit more complex than just blowing it into the cells.

Actually there are many problems to solve,

1: creating metal particles small enough to penetrate the nucleus of a cell without destroying it (100 nm?)
2: loading the dust onto a clean surface
3: hitting that surface hard enough to eject the dust into the plant material in a but soft enough not to destroy the material or the material below it
4: building a stage for doing it at the same height every time.

All of these problems have solutions.  I think a DIY gene gun would get a lot of media play, however, I don't think it would make genetic engineering any easier to do. For example, RNA interference can be done with a syringe, and Agrobacterium can be applied just by soaking the tissue. Callus culture on antibiotics is the gold standard, but you can do photobleaching markers instead. Gene guns require tissue culture 100% of the time, and I'm not sure they have any better efficiency (last I checked for Agrobacterium in Capsicum you got about 1 / 1000 calluses that make it to genetically modified plants).

Ben

[Conner Berthold]

[Andreas Sturm]

The pdf sounds great. Just printed it out ;)

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[Conner Berthold]

I was also looking to see if Helium could be replaced with something more "portable" (like CO2 cartridges) and found this article.

"With the self-design CO2 propelled gene gun (200 

psi, distance 3 cm with 400 mesh nylon screen) using 

tungsten particle (600 nm diameter) coated with plasmid 

expressing anti-ampicillin gene, the plasmid with antiampicillin gene was high efficiently transferred into E. 

Coli cells."(http://www.sciencepub.net/nature/0204-supplement/03-mahongbao-schistosome.pdf)(3rd page)

This link also has an interesting approach to prevent harm by preventing the shock wave of gas from directly hitting the culture. If you could get both of these to work a smaller portable gun should be possible.

[Cathal Garvey]

Helium is used because it can achieve higher speeds, but you don't need
that much power to punch through a bit of peptidoglycan/cellulose and a
lipid bilayer. :)

On 28/10/12 19:21, Conner Berthold wrote:
> I was also looking to see if Helium could be replaced with something more
> "portable" (like CO2 cartridges) and found this article.
>
> "With the self-design CO2 propelled gene gun (200
> psi, distance 3 cm with 400 mesh nylon screen) using
> tungsten particle (600 nm diameter) coated with plasmid
> expressing anti-ampicillin gene, the plasmid with antiampicillin gene was
> high efficiently transferred into E.
> Coli cells."(
> http://www.sciencepub.net/nature/0204-supplement/03-mahongbao-schistosome.pdf)(3rd
> page)
>
>

> This link <http://physics.ucsd.edu/~groisman/Gene%20guns.html> also has an

> interesting approach to prevent harm by preventing the shock wave of gas
> from directly hitting the culture. If you could get both of these to work a
> smaller portable gun should be possible.
>

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[Andreas Sturm]

Sounds quite doable, because we have a compressor at home.


But where can you place the coated particles? I'd close the 'tube' with Parchment paper on the downside. On the top there is the compressor attached, with a electrical valve to quickly release the pressure.

Will the parchment paper allow the tungsten particles to get through, without leaving the DNA on the paper?

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[Patrik D'haeseleer]

Here's some nice videos on Jove, on preparation of gene "bullets", and use of gene gun technology to transfect neuronal and plant tissues. Highly recommended:

http://www.jove.com/video/675/preparation-gene-gun-bullets-biolistic-transfection-neurons-slice

http://www.jove.com/video/1963/bimolecular-fluorescence-complementation-bifc-assay-for-protein

Making the gene "bullets" yourself looks entirely feasible, given the right cobbled-together equipment...

Patrik