Simultaneous RNA and DNA transfection? - (Sep/11/2013 )
I want to transfect RNA (viral) with a reporter gene into huh7 cells. I need to normalize the transfection efficiency of the RNA to another reporter gene that is in a DNA plasmid with its own promoter. Is it possible to mix the RNA and DNA for a single transfection? I can only find examples of 1 or the other, not both. Any advice would be greatly appreciated.
We cotransfect short RNA (siRNA or miRNA) with DNA plasmid without a problem. Probably you need to figure out the right ratio between RNA and DNA since they compete for transfection reagent, e.g., lipofectamine.
I have found protocols for miRNA and siRNA. I however will be using a 7kb capped RNA with a DNA plasmid that will be around 4-5kb. Maybe I should add a large excess of lipofectamine since they will compete for the reagent?
To remove variables, I wanted to mix the rna and dna and then conjugate with lipofectamine. The only other option I came up with was to make rna transcripts from the dna control plasmid. But this is not that easy since the commercially available plasmids don't have a encoded polyA tail but use the SV40 polyA signal.