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ShRNA Screening and Histopathology

Subject Area Hematology, Oncology
Term from 2015 to 2023
Project identifier Deutsche Forschungsgemeinschaft (DFG) - Project number 267467939
 
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.
DFG Programme Research Units
 
 

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