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Inducing RNAi with siRNA Cocktails Generated by RNase III

of generating a population of siRNAs that can effectively silence target genes. The siRNA population generated by this method does not exhibit higher levels of toxicity (data not shown) or nonspecific effects on gene expression as compared to individual chemically synthesized siRNAs. The use of RNase III to prepare siRNA cocktails eliminates the need for expensive chemical synthesis, laborious siRNA expression vector production, or the time-consuming screening necessary to identify an effective siRNA sequence. RNase III generated siRNA cocktails are a good alternative to standard methods of siRNA production for the researcher that needs a quick result and does not require siRNA sequence information.

To simplify the preparation of siRNAs by RNase III digestion, Ambion now provides the Silencer siRNA Cocktail Kit (RNase III). This kit includes reagents and detailed protocols for generating and purifying dsRNA, dsRNA digestion by RNase III, and subsequent siRNA clean up. For more information, see the article Avoid siRNA Design Altogether.

Figure 3. RNase III siRNA Cocktails Show Specificity for Silencing. HeLa cells were transfected with 100 nM RNase III generated siRNAs to GAPDH. Immunofluorescence analysis of GAPDH, La, c-MYC, Cdk-2, Ku-90, and -actin was performed 48 hours post transfection and subsequently quantitated.


Materials & Methods
Preparation of siRNA Cocktails with RNase III Total RN
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