8
$\begingroup$

I see a lot of folks using different techniques for transforming mammalian cell (specifically HEK) instead of doing electroporation like I see with E.Coli (bacteria). Is there a reason for this ?

$\endgroup$

1 Answer 1

8
$\begingroup$

Viral methods are just more efficient and give you a higher viability in the end. Electroporation works but you'll have a low yield of viable cells after. Here's a nice paper looking at the effects of buffer composition on cell viability/transfection efficiency: https://www.nature.com/articles/s41598-020-59790-x with the general take away being, if electroporation is working well enough to transfect the cells, it will also kill a fair percentage as well.

Hek cells in particular are pretty easy to transfect through viral methods for stable integration of your plasmid or Fugene/lipofectamine for transient expression.

$\endgroup$
7
  • $\begingroup$ The reason mild electroporation has low transformational efficiency in eukaryotic cells is because they sequester their DNA behind a nuclear membrane, right? $\endgroup$
    – timeskull
    Commented Mar 16, 2021 at 14:53
  • $\begingroup$ I'm pretty sure that all transfection techniques like electroporation/CaCl2/Lipofectamine are designed just to break the hurdle of getting it into the cytoplasm not the nucleus. The nucleus is a barrier to transfection but the prior mentioned methods aren't responsible for getting it there. Another shout out to retroviral based methods which will bring it to the DNA for you and integrate it into the genome $\endgroup$
    – yp66t89
    Commented Mar 16, 2021 at 20:18
  • $\begingroup$ @yp66t89 I guess my followup question would be: "Why don't people use the retroviral methods for bacteria ?" Is there something that makes it not work, or is it just that electroporation would just be cheaper. $\endgroup$ Commented Mar 17, 2021 at 14:59
  • 1
    $\begingroup$ I'd assume bacteria tend to be more efficient at both integrating plasmids than mammalian cells, no one cares if most of your E.coli die as long as one bacterium survives with your plasmid, and I don't actually know if you can use virus on bacteria, I think you'd need a bacteriophage $\endgroup$
    – yp66t89
    Commented Mar 17, 2021 at 18:02
  • $\begingroup$ @yp66t89 A bacterialphage is a virus --- that's the name for a virus that infects bacteria. $\endgroup$
    – jakebeal
    Commented Mar 19, 2021 at 8:42

You must log in to answer this question.

Not the answer you're looking for? Browse other questions tagged .