CCR RNAi initiative: Establishment of shRNA RNAi library

在2005年期间，我们已经建立了一些关键的参数所需的利用小RNA文库的大小，以整个基因组。该项目遇到了许多技术问题，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资源的显着改进鼓励我们购买整个人类基因组预构建的shRNA文库（10万个克隆）。我们现在已经标准化了细菌储备的生长条件，并正在最终确定与高通量规模兼容的纯化方法。我们已经确定了我们最有可能的原理验证筛选，最初使用库的优先子集来研究人乳头瘤病毒（HPV）感染及其在人宫颈癌中的作用。

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.