Can the method of electroporation be coupled with viral vector gene transfers?
I haven't come across any papers that talk about it?
Kindly suggest if anyone is aware of anything helpful.
Very often in transfection techniques, the use of two different transfection systems may work equally as well when used alone, but become less efficient in combination. This appears to be true with gene transfer by infection and electroporation used together. See the following paper:
http://drugdelivery.chbe.gatech.edu/Papers/2003/Coulberson%20J%20Controlled%20Release%202003.pdf
You can ask it from my dear Professor via https://www.linkedin.com/in/ali-afgar-982374139/
Hi Pranav,
I have done this in the past but sequentially. Meaning I used electroporation to deliver cre/lox reporters and some days later I added AAV-Cre viruses. These experiments were done in cultures of primary embryonic neurons, which are quite sensitive, and it worked very well.
I assume you can also perform shorter protocols, i.e. you can electroporate de cells and infect already one day later if the cells are healthy and properly attached.
Let me know if you need more details.
Best.
Sequential use would be the experimental design that made the best use of both technologies in a single experiement, Sebastián Dupraz !
Sebastian , BUT would the same sequential manner work in vivo?
Thats the big queston for me.
Cheers
Hi Pranav,
I never tried these experiments in vivo. However, if using this sequence you manage to target the same cells (meaning cells do not migrate/relocate after the initial electroporation) it may work like in vitro. For instance, I would not use it for a sequential infection after a couple of days passed an brain in utero electroporation since the the neurons electroporated would migrate in the cortex and wouldn't be efficiently targeted by infection of progenitors in the subventricular zone of the brain. Here the approach would change to target the transfected cells with the virus their new "current" location (i.e. using stereotaxic injection of virus).
May I ask which cells/tissue are you planning to use?
Best,
Ok thanks a lot Sebastian .
I am thinking of trying this technique out in retina tissue and am not really sure incase electroporation would work well with viral vectors in general.
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