Easing siRNA Transfection

Image: Courtesy of Ambion SILENCE! HeLa cells transfected with Cy3-labeled GAPDH or Scrambled siRNAs using Ambion's siPORT Lipid Transfection Agent were harvested 48 hours post-transfection. Red: Cy3-labeled siRNA; blue: DAPI-stained nuclei; green: GAPDH protein. Introduction of the scrambled control siRNA (left) had no effect on GAPDH protein expression, whereas transfection of an siRNA targeting GAPDH (right) resulted in drastic reduction in GAPDH protein levels. Much attention has bee

Jeffrey Perkel

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Much attention has been paid recently to RNA interference (RNAi), a technique in which exogenous, double-stranded RNAs (dsRNAs) are introduced into a cell to specifically destroy a particular mRNA, thereby diminishing or abolishing gene expression.¹ The technique has proven effective in Drosophila, Caenorhabditis elegans, plants, and recently, in mammalian cell cultures.²

To make the technique work in cultured mammalian cells, researchers must deliver small interfering RNAs (siRNAs), which are dsRNAs of 21-25 nucleotides, to the cell. They can use a wide range of standard transfection reagents optimized for DNA and RNA, but according to Kathy Latham, RNAi product manager for Austin, Texas-based Ambion, reagents designed to deliver mRNAs, for instance, are not adept at transfecting smaller siRNAs. "It's like the difference between plasmids and oligos," she says, referring to manufacturers' offering different formulations to transfect plasmid DNA and shorter oligonucleotides.

As a result, both Ambion and Madison, Wis.-based Novagen have ...

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