Transient transfection on a stable cell line - (Feb/05/2006 )

Hi everyone!

I have got a stable transformed cell line. I was wondering if I can do transient transfection with another plasmid. Will I get a transient transfection of the two proteins? Those two proteins are supposed to assemble together. Is this a proper scientific methodology?

I would like to get your comments, suggestions and opinions.

Thanks

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hi
you can do a first try with a reporter protein (GFP, DsRed...) and you'll see if transient transfection occurs.
I can say, that a colleague, has stablyintegrated 5 different proteins in the same cell and all 5 are expressing themselves correctly.
So i don't think that will be a problem.

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I've done this, it's no problem at all! (cells I've done this on: U87 already transfected with 2 extra (selected) proteins and Hela cells already transfected with 2 proteins and 2 other sequences, thus a total of 4 different pieces of separate DNA integrated stably).

But as Fred suggests: first try it with a reporter protein, just to make 100% sure.

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Thanks. Yes, I will do as Fred said with the reporter sequence. Now I'm confident it will work.

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I also want to co-tranform two plasmids in one cell.
Now I know, Thanks!

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The answer is a little late maybe but I have done that also... I used CHO cells stable transfected with a receptor and transfected them temporarely with different protein.

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technically spoken there should be no limits to generate multi-gene stable transfectants;

however, the biology is the more important question which means if single or multiple transfected genes are really co-expressed or even if expressed, do damage the cell

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