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Assessing Gene Function with siRNA Libraries

libraries of effective siRNAs. This particular data set allows the researcher to go beyond a primary readout (affects on cell proliferation in this case) to efficiently and directly address underlying causes such as cell cycle deregulation at several levels with a single screening experiment. By comparing the cell proliferation and mitotic index data, one can relate the antiproliferative effects observed for several kinase-targeting siRNAs back to underlying effects on cell cycle regulation.

Silencing kinases necessary for passage of the cell through G1, S, and G2 phases of the cell cycle may be expected to lengthen interphase and therefore decrease the percentage of cells in mitosis. It will also decrease the overall cell proliferation rate. Several of the siRNAs investigated here induced this phenotype (Figure 4). Similarly, inhibition of kinases required for passage through mitosis would be expected to cause an increase in mitotic index and a decrease in cell proliferation. Several siRNAs also appeared to induce this phenotype.

Figure 4. Deviations in Cell Number and Mitotic Index Induced by siRNAs Targeting 178 Kinases. Plot of the data from Figures 2 vs. Figure 3 demonstrating that silencing of certain kinases dramaticly affects the cell cycle. Multiple categories of data can be defined, such as 1) no change, 2) increase in cell growth with no change in mitotic index, 3) no change in cell growth, increase in mitotic index, etc. Shown in red are cells exhibiting arrest in G1, S or G2 (left), and cells exhibiting arrest in M (right), phases of the cell cycle.
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