| Project Overview: | The S phase kinase Cdc7 plays an essential role in the initiation of DNA synthesis where it activates the pre-initiation complex by phosphorylating key components of the complex, for example MCM2, a component of the hexameric helicase required for unwinding the DNA. SiRNA depletion in cancer cell lines has led to cell death whereas the depletion has resulted in G1/S arrest in non-transformed cell lines. Thus, giving raise to the proposal that inhibition of Cdc7 would cause tumour specific cell kill. Treatment of cancer cell lines with Cdc7 specific small molecule inhibitors have resulted in cell death, however, this is not seen in all cancer cell lines tested. Thus, we would like to screen for synthetic lethal targets which could be used to help identify the patient population with which to treat with the Cdc7 inhibits to achieve the best therapeutic effect in the clinic. The screen will be carried out with 2 different cell lines, MCF7 and A549 using the Cdc7 inhibitor CRT0274128 and the siRNA whole genome library using cell count as the assay read-out. The cells will be transfected with the siRNA library and treated with the Cdc7 inhibitor 24hr later and assayed 72hr after inhibitor addition. The screen will be done in triplicate. The Cdc7 inhibitor will be used at a fixed concentration which is able to elicit an on target inhibition of the MCM2 phosphorylation, a direct substrate of the Cdc7 kinase with minimal phenotype effect to ensure a signal window for the screen. |
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| Screener: | Flo Chan (CRT Lab). Extension:CRT, Rm CRT |
| Project Status: | Primary Screen |
| Project Start Date: | Mar 2012 |
| Keywords: | Cdc7 inhibitor Viability |
| Libraries Screened: |
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