White Paper: Efficient Transfection of shRNA-encoding Plasmids into Mammalian Neurons

Transfection methods are widely used to study miscellaneous aspects of cell biology. The transfection of plasmids encoding tagged proteins, for instance, allows the visualization of the location and behavior of proteins in living cells. With the discovery of RNA interference (RNAi), it is now also possible to specifically prevent the production of certain proteins within a cell via the transfection of plasmids encoding shorthairpin RNAs (shRNAs). Transfection of shRNA-encoding expression plasmids has several advantages over a transfection with siRNAs. For one, it ensures the knock-down of the targeted protein over longer periods of time via the prolonged expression of the shRNA. Moreover, by including inducible elements in the promoter, the expression of an shRNA can be switched on and off as desired. Finally, constructs can be designed for the stable integration of the shRNA-encoding sequence into the genome. As a consequence, such approaches will greatly accelerate our understanding of gene function in a cellular context.