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Are siRNA Pools Smart?

in apoptosis (Figure 4). Consistent with the cell proliferation results, the single siRNAs for each gene had similar, though non-identical effects, on caspase 3 activity. Also consistent with the cell proliferation results, the siRNA pools only partially overlapped the results of the single siRNAs (Figure 5). Examples of single and pooled siRNA results are shown in Figure 4B.

Figure 5. Overall Performance of Single siRNAs and siRNA Pools. Using the definition of a hit as a gene for which two single siRNAs produce a phenotype that is significantly different than samples transfected with a negative control siRNA, the performance of each of the single siRNAs (siRNA #1, #2, or #3) as well as the siRNA pool of the three siRNAs was compared. The combined results of screening with two (#1+#2, #1+#3, and #2+#3) or three (#1+#2+#3) individual siRNAs illustrates the higher accuracy of this strategy. Accurate predictions of hits and inaccurate predictions of hits are shown in green and orange, respectively. False negatives are shown in grey.


Single siRNAs vs. siRNA Pools: Results

To minimize experimental variability, the data comparing the effects of single and pooled siRNAs on proliferation and caspase 3 activation were generated with the same cell line and substrates at the same time. The results of the screens were used to determine the effectiveness of transfecting a series of three single siRNAs versus transfecting a pool of the three siRNAs for identifying gene "hits." The definition of a hit in our screen was a gene for which two single siRNAs gener
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