CCR RNAi initiative: Establishment of shRNA RNAi library

在2005年，我们已经建立了一些关键参数所需的shRNA文库在整个基因组的大小利用。这个项目遇到了一些技术问题，其中许多与shrna的内在不稳定性有关。然而，最近由校外社区开发的shRNA资源的重大改进鼓励我们购买全人类基因组预构建的shRNA文库（100,000个克隆）。我们现在有了标准化的细菌存量生长条件，并正在最终确定与规模化高通量兼容的纯化方法。我们已经确定了我们最可能的原理验证筛选，最初使用文库的优先子集来研究人类乳头瘤病毒（HPV）感染及其在人类宫颈癌中的作用。

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.