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Optimizing Chemical Transfection and Electroporation of siRNAs

nt, but multiple pulses may be required to achieve the desired level of silencing with siRNA in some cell types. Because the fine-tuning of electroporation involves varying parameters that can significantly influence cell viability, for these optimization experiments, studies to determine cell viability before and after electroporation events can be crucial.

Transfection efficiency is also affected by the concentration, the purity, and the size of the nucleic acid used. As with chemical methods, transfection efficiency can be modulated by varying the siRNA concentration within a limited concentration range. Experiments with different cell types reveal that the effective concentration of siRNA is somewhere between 100 nM to 1 M in a standard electroporation reaction. siRNA amounts in excess of 2 M may lead to an increase in the transfection efficiency, but may also contribute to cytotoxicity in some cell types. (Experiments detailing these parameters are provided in the article Efficient Delivery of siRNAs to Human Primary Cells.) Although transfection efficiency is affected by the size of the nucleic acid electroporated, we have found that parameters optimized for plasmids can usually be used as starting points for optimizing electroporation of siRNA.


siRNA Experiments Addressing Electroporation Variables
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