I am trying to transfect HEK cells with 15-20 different plasmids and it seems there is some kind of competition for expression. I am currently using Lipofectamine 2000 and transfect each plasmid in 1 ug concentration.
Whether you are transfecting one plasmid or twenty isn't really as important as how much total DNA you are transfecting. I've easily transfected these cell lines with 14 ug DNA when plated at 4.6 million cells on a 10 cm dish. I think as long as you can adjust your plasmid concentrations such that the total DNA:lipid regent doesn't kill the cells, you'll be fine. We've used LF3000 we well as several of the Mirus reagents with great success.
A bigger concern though would be do all of the plasmids get in each cell in a population. I think most cells will get most plasmids, but there will likely be a bit of variation. As long as you have enough replicates to account for this, I would think you'd be golden.
Hi Lena,
Baculovirus multigene delivery it's an option, check out this paper from my group.
https://www.nature.com/articles/ncomms11529
Hope it helps
Francesco
Hi Lena,
I think that is might be challenging to transfect 15-20 plasmids at once and expect a consistent expression of all your constucts:
1) are they all about the same size?
2) are they all on the backbone vector/promotor?
3) have you considered an alternative strategy?
Thanks,
Nicolas
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