Transient transfection on a stable cell line - (Nov/04/2006 )

Hi all,

This is my first time posting and I hope to get some help from all of you.

Transient transfection Gene A + B = Effect

Transient transfection Gene B into stable cell line with Gene A = No Effect

G418 selection for the stable cell line.

Is there any reason for this?

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We had in the past the problem that a stably transfected cell line became resistent to transfectio: maybe this could be your case as well. Why don't you try to transfect your stable cell line with a GFP expression plasmid and have a look at whether they are transfectable or not?

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Maybe for your specific cell line this might be a problem, but I've performed transfections on cells lines that were stably transfected with 2 different genes (puromycin and geneticin selection), was no problem at all.

So I would try transfection with GFP (or other easily detectable genes).

What's the "effect" you're talking about, any reason why maybe gene A might no longer be active and hence you get no effect? (do you check expression of gene A, things can change after some passages...).

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