( Dmitriy Ovchar...)Efficient Delivery of siRNAs to Human ,,, Primary Cells: Electroporation ,,, vs. Chemical Transfection

Dmitriy Ovcharenko (Research Associate II, Ambion, Inc.)

Gene silencing using small interfering RNAs (siRNAs) has become a powerful method for studying gene function. Human primary cells are often desired for such experiments because they are more similar to their in vivo counterparts than are immortalized cells. The use of siRNAs in human primary cells continues to accelerate applications such as target validation, gene discovery, and gene therapeutic approaches that could lead to powerful new gene-specific siRNA-based therapeutics.

Chemical transfection is the standard method for introducing siRNA into immortalized cells. Unfortunately, efficient transfer of siRNAs into primary cells by chemical transfection has so far been restricted to a few cell types only. This is illustrated in Figure 1, below. Primary cells tend to be more difficult to transfect chemically in general, which has limited their use for siRNA experiments. Here we address the use of electroporation as an alternative for efficient delivery of siRNAs into primary cell types.

Chemical Transfection: Inefficient siRNA Delivery in Primary Cells
To show that chemical transfection is only useful as an siRNA delivery method for only certain types of primary cells, an siRNA targeting GAPDH was transfected into normal human umbilical vein endothelial cells (HUVEC) and normal human epidermal
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