RNA Interference Tools Core

核心B的总体目标是在涉及基于RNAi的拆卸的所有方面支持此POI 技术参与项目在很大程度上依赖于蜂窝系统作为发现的平台， 以及我们评估基因功能的能力。在缺乏基因遗传方法的情况下 在体细胞中进行消融，RNAi是一种快速、高效、廉价、 通用、易用的基因敲除方法。尽管RNAi方法也提供了一些 为应对这些挑战，核心小组将制定应对挑战的工作战略。我们的首要目标是 产生多种高效的RNAi试剂来抑制我们期望的靶点的表达 在这种PPG过程中出现，作为PTEN-Siah2途径的潜在调节因子。这些试剂将是 有助于评估这种调节剂作为治疗靶点的潜力 肿瘤类型源于PTEN缺乏症。为了实现这一目标，我们将系统地生成多个 慢病毒shRNA，并从经验上评估它们的击倒效力。选定的试剂将是 被转移到各种细胞系，并在该计划的不同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一种公正的策略。 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、专业知识和人力来开发高吞吐量 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，它将向这些项目提供 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.