why the production of a protein by cells transfected with siRNA (in-vitro) is less than the production of that protein by untransfected cells?
Thanks in advance
Sarah:
Theoretically it shouldn't be different but during the transfection with negative control siRNA along with transfection reagents like Lipofectamine 2000 or RNAimax, a significant cell death occurs which substantially affect the results in comparison to the untransfected control cells. Cell death is basically due to the transfection reagents which are toxic to cells. Such cell death or cytotoxicity leads to the false positive results.
Hope this helps!
No it is not the lipofectamine because I tested it using other siRNA of a specific gene and the amount of protein produced was the same as the untransfected.
My doctor once told about binding of scramble siRNA to nonspecific target but it was not clear for me when I googled it
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