( Rich Jarvis (As...)Optimizing Chemical Transfection and Electroporation of siRNAs

Rich Jarvis (Associate Scientist, Ambion, Inc.)

Alan D. King (Vice President and CSO, Cyto Pulse Sciences, Inc.)

Low transfection efficiency is the most frequent cause of unsuccessful gene silencing experiments. We have found that by optimizing a few critical variables, higher levels of transfection efficiency can be achieved in many cell types.

Good Transfection is Critical
The ability of small interfering RNAs (siRNAs) to silence gene expression is proving to be invaluable for studying gene function in cultured mammalian cells. siRNAs or DNA constructs that express siRNAs can be transiently transfected into mammalian cells using standard techniques. However, obtaining efficient siRNA delivery by chemical transfection or electroporation is not trivial and may limit the utility of RNAi in some cell types.

A variety of methods have been developed for transfecting nucleic acids into cultured cells. The choice between chemical transfection or electroporation is largely dependent on the cell type and the characteristics of the nucleic acid being transfected. Some cell types, including most immortalized cell lines, can be transfected with any number of commercially available chemical transfection agents with careful optimization. Other cell types, such as some finite cell l
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