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Assessing Gene Function with siRNA Libraries

ure 2. Changes in Cell Proliferation Induced by 178 Kinase siRNAs from the Silencer Kinase siRNA Library. HeLa cells were transfected with individual siRNAs targeting 178 different kinases as described in Figure 1. Forty-eight hours post-transfection cells were counted. Cell numbers per microscopic image are given.


Changes in Mitotic Index
One way to better understand a gene's role in cell cycle progression is to monitor the percent of cells undergoing mitosis at any given point in time, with and without siRNA treatment. For asynchronously growing cultures such as those used in these experiments, the mitotic index reflects the fraction of time that cells spend in mitosis versus the rest of the cell cycle. Thus, as shown in Figure 3, the ~2.5% mitotic index value observed for control cells transfected with a scrambled siRNA indicates that cells spent 2.5% of the cell cycle in mitosis.

Figure 3. Mitotic Index of Cells Transfected with 178 siRNAs from the Silencer Kinase siRNA Library. HeLa cells were transfected with individual siRNAs targeting 178 different kinases as described in Figure 1. Forty-eight hours post-transfection mitotic index was measured. The orange bar shows the percent of negative control cells, which were transfected with a control scrambled siRNA, undergoing mitosis (mitotic index range of 2.1 to 2.3). The semitransparent brown rectangles highlight the 95th percentile range of normal mitotic index. The inset shows two interesting mitotic arrest phenotypes induced by siRNAs targeting two different kinases (green = chromatin; orange = tubulin).


Several siRNAs induced dramatic changes in the mitotic index wh
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