I am trying to express my HA-tagged viral protein in S2 cells. This protein is known to inhibits the cellular transcription. Therefore I tried to transfect with invitro transcribed capped and polyadenylated RNA. However, I am not getting the expression. I tried transfection with different Lipofectamine RNA ratios, time points and also with different batches of RNA. Still, I couldn't solve the problem. Any suggestions?
I would suggest to test first mRNA transfection with a control, e.g. GFP encoding mRNA or a labeled mRNA. To be sure that you have a cell uptake.
https://viromer-transfection.com/product/plasmid-transfection-control-viromer-red-gfp
Hi try using Lipofectamine 3000 (which works better in some cells vs 2000) and use positive and neg controls. inefficient delivery could be one problem, but mRNA could be another problem
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