old protocol - transformation without injection?

Model Organism: Worms Methods & Reagents
1

Hi all,

I’m looking for an old report from the mid/late 90s that described transgenic transformation without microinjection. The protocol involved abrasive microfibers, DNA, worms, and vortexing (I know, it sounded just as crazy then as it does now). My online searches have been unsuccessful, so I’m hoping that someone in the community will come to my rescue.

Thanks,
Harold

2

Was it this one?

Genetic transformation of nematodes using arrays of micromechanical piercing structures.
Hashmi S, Ling P, Hashmi G, Reed M, Gaugler R, Trimmer W.
Biotechniques. 1995 Nov;19(5):766-70.

3

I’ve seen another variant of the report Barth Grant highlights (same method, same authors, different species).

In other transformation without microinjection (and without expensive microparticle bombardment apparatus): electroporation is reported to work, albeit inefficiently.

4

Thanks, Barth and Hillel, but the protocol I’m seeking did not utilize a microfabricated device. My (hazy) recollection is that the worms are added to a solution containing transgene plus abrasive fibers, then vortexed to generate micropunctures that introduce the transgene. But it’s conceptually similar to the reference you provided, so I may be able to use the dimensions of their microprobe to track down some appropriate materials to test.

-Harold

P.S. - Re: electroporation, I tried it (long ago), but only managed to create worm-worm fusions at high voltage. Not pretty…

5

I haven’t found anything with nematodes, but there seems to be some literature for the transformation of plants and yeast by agitation with “silicon carbide whiskers”.

6

Bingo! “Whiskers” was the term I was looking for. I’ll follow up with the worm reference if/when I find it.

Thanks for your persistence, Hillel!

-Harold