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Silencer Validated siRNAs

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Figure 1. Overall Performance of siRNAs Targeting Sixty Genes. siRNAs targeting 60 human genes were designed using the algorithm developed by Cenix. The top siRNA candidate for each target was prepared and transfected. Forty eight hours post-transfection, target gene expression was quantified by real-time RT-PCR. Relative reduction in mRNA expression was measured against cells transfected with a negative control siRNA. Sample size was normalized by measuring GAPDH in the various samples using real-time PCR.


Characteristics of Silencer Validated siRNAs
The average reduction in target gene expression achieved with the inital 45 Silencer Validated siRNAs tested is greater than 90% as measured by real-time RT-PCR, indicating that the design algorithm used is not only able to identify effective siRNAs at a high rate, but is also able to identify highly effective siRNAs. To further characterize the initial set of validated siRNAs, the potencies of a handful of the siRNAs were measured after transfecting HeLa cells with siRNAs at five different concentrations, ranging from 1-100 nM. As seen in Figure 2, each of the siRNAs provided a similar level of mRNA knockdown in cells transfected with 10 nM, 30 nM, and 100 nM siRNA. This confirms the potency of the siRNAs designed using the algorithm developed by Cenix, as well as the quality of the synthesis and purification processes being employed by Ambion for the Silencer Validated siRNAs. The siRNAs targeting survivin, CDK2, and GAPDH were tested further by transfecting HepG2, SK-N-AS, and MCF7 cells with five different concentrations of the siRNAs. Even in these more difficult to transfect cells, the Silencer Vali
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