During 2005 we have established some key parameters required for the utilization of shRNA libraries at a size up to whole genome. This project has encountered a number of technical problems, many relating to the intrinsic instability of shRNAs. However, recent significant improvements in shRNA resources developed by the extramural community have encouraged us to purchase a whole human genome pre-constructed shRNA library (100,000 clones). We now have standardized growth conditions for the bacterial stocks and are finalizing purification methods compatible with scaling to high throughput. We have identified our most likely proof-of-principle screen to be conducted using initially a prioritized sub-set of the library to study human papilloma virus (HPV) infection and its role in human cervical cancer.
