RNA Interference Tools Core

核心B的总体目标是在涉及基于RNAi的敲低的所有方面支持该POI 技术.参与的项目在很大程度上依赖于蜂窝系统作为发现的平台， 以及我们评估它们基因功能的能力。在缺乏基因遗传学方法的情况下， 在体细胞中的消融，RNAi是用于快速、有效、廉价， 通用且易于使用的基因敲除方法。尽管RNAi方法也存在一些问题， 面对这些挑战，核心将制定工作战略，以克服这些挑战。我们的首要目标是 产生多种高效的RNAi试剂来抑制我们预期将 作为PTEN-Siah 2途径的潜在调节剂在PPG过程中出现。这些试剂将 有助于评估这种调节剂作为治疗靶点的潜力， 癌症类型源于PTEN缺陷。为了实现这一目标，我们将系统地生成多个 慢病毒-shRNA并凭经验评估其敲低效力。所选试剂将 转导到各种细胞系中，并在该计划的各个阶段转移到项目团队。 我们的第二个重点将是siRNA筛选，作为发现新的生物学环境的无偏见策略。 PTEN相关的肿瘤发生和精确定位新的潜在靶点。为了实现这些目标，我们将 提供RNAi文库，最先进的设备，专业知识和人力，以开发高通量 基于细胞的分析，执行筛选，分析和确认结果，并生产优化的试剂， 后续研究。核心B将与3个项目和核心C密切互动， 试剂和服务。

The overall goal of Core B is to support this POI in all aspects involving RNAi-based knock-down technology. The participating Projects rely to a great extent on cellular systems as platforms for discovery, as well as on our ability to assess gene-function in them. In the absence of genetic approaches for gene ablation in somatic cells, RNAi is the ideal tool for such purpose as a rapid, efficient, inexpensive, versatile, and easy to use method for gene knock-down. Although RNAi approaches also present some challenges, the Core will put in place working strategies to overcome them. Our first aim will be to generate multiple, high-efficiency RNAi-reagents to inhibit the expression of targets that we expect will arise in the course of this PPG as potential regulators of the PTEN-Siah2 pathway. These reagents will be instrumental in evaluating the potential of such regulators as therapeutic targets for the treatment of cancer types derived from PTEN deficiency. To achieve this goal we will systematically generate multiple lentiviral-shRNAs and empirically assess their knock-down potency. The selected reagents will be transduced into the various cell lines and transferred to project teams at various stages of this program. Our second focus will be siRNA screening, as an unbiased tactic to discover new biology surrounding PTEN related tumorogenesis and pinpoint new potential targets. To accomplish these goals, we will provide RNAi libraries, state of the art equipment, expertise and manpower to develop high throughput cell-based assays, execute screens, analyze and confirm results, and produce optimized reagents for follow-up studies. Core B will interact closely with the 3 Projects and Core C, to which it will be providing reagents and services.