ShRNA Screening and Histopathology

ShRNA 筛选和组织病理学

• 批准号: 280454646
• 负责人: Professor Dr. Martin Eilers
• 金额: --
• 依托单位: Pathologisches Institut
• 依托单位国家: 德国
• 项目类别: Research Units
• 财政年份: 2015
• 资助国家: 德国
• 起止时间: 2014-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/280454646?language=en
• 关键词: ShRNA; Screening; Histopathology

This central project provides critical technologies to this research consortium. Specifically, Z01 has three tasks:First, this project provides the synthesis of focused shRNA libraries and and the analysis of screens with these libraries. In particular, the project provides focused shRNA libraries as well as technical advice for both in vitro and in vivo shRNA screens, advises individual projects on set-up of in vivo experiments (e.g. animal number), and recovers and sequences the genomic DNA containing the retroviral inserts. The project also analyses the sequencing data including a statistical analysis. As part of this service, the Z01 also handles all communication with Mirimus and the handling of shRNA-transgenic mice that are imported into the consortium. The second task of this project is the target validation using a pipeline of established stress assays. This part involves the analysis of multiple stress parameters after depletion of specific targets or interference with small molecules using high-content microscopy and optical image analysis. These assays are established and data are provided as quantitative evaluation to all members of this consortium. This screening core facility will also perform small molecule screens of FDA-approved compounds in the next funding period. The third task of this project is to provide mouse and human pathology analyses with the aim to validate therapeutic targets (e.g. by analyzing their expression in human tumors) and to study the in vivo effects of interfering with essential processes in tumors. In the next funding period, this part of the project will include a standardized analysis of immune cell infiltrates into tumors after therapeutic intervention. This analysis pipeline and the necessary staining protocols have been established.

这个中央项目为该研究联盟提供了关键的技术。具体而言，Z01具有三个任务：首先，该项目提供了聚焦的ShRNA库的综合以及使用这些库对屏幕进行分析。特别是，该项目为体外和体内shRNA筛查提供了重点的shRNA图书馆以及技术建议，并向单个项目提供了有关设置体内实验（例如动物数量）的建议，并恢复并序列包含逆转录病毒插入物的基因组DNA。该项目还分析了包括统计分析的测序数据。作为这项服务的一部分，Z01还处理了与Mirimus的所有通信以及进口到财团的shRNA-转基因小鼠的处理。该项目的第二个任务是使用已建立的压力测定管道的目标验证。该部分涉及对特定靶标的耗尽后多应力参数的分析，或使用高含量显微镜和光学图像分析干扰小分子。建立这些测定法，并向该财团的所有成员提供数据作为定量评估。该筛选核心设施还将在下一个资金期间执行FDA批准化合物的小分子筛选。该项目的第三个任务是提供小鼠和人类病理分析，目的是验证治疗靶标（例如，通过分析其在人肿瘤中的表达），并研究干扰肿瘤基本过程的体内影响。在下一个资金期间，该项目的这一部分将包括对治疗干预后免疫细胞浸润到肿瘤中的标准化分析。该分析管道和必要的染色方案已建立。