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Simultaneous RNA and DNA transfection?


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#1 HOYAJM

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Posted 11 September 2013 - 08:27 PM

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.

 

Thanks



#2 pcrman

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Posted 15 September 2013 - 05:35 PM

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.



#3 HOYAJM

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Posted 15 September 2013 - 06:05 PM

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.




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