CCR RNAi initiative: Establishment of shRNA RNAi library

CCR RNAi倡议：建立shRNA RNAi文库

• 批准号: 7292896
• 负责人: natasha caplen
• 金额: --
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7292896
• 关键词: CCR; RNAi; initiative; Establishment; shRNA

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.

在2005年期间，我们已经建立了一些关键参数，以便在整个基因组范围内利用shRNA文库。该项目遇到了一些技术问题，其中许多与shRNAs的内在不稳定性有关。然而，最近由校外社区开发的shRNA资源的显著改进鼓励我们购买预先构建的全人类基因组shRNA文库(100,000个克隆)。我们现在已经有了标准化的菌种生长条件，并正在敲定与规模化到高通量兼容的纯化方法。我们已经确定了我们最有可能进行的原则证明筛查，最初使用文库中优先考虑的子集来研究人乳头瘤病毒(HPV)感染及其在人类宫颈癌中的作用。