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Systemic shRNA screens and high throughout sequencing to investigate pathway dependence in multiple myeloma

Subject Area Hematology, Oncology
Term from 2009 to 2014
Project identifier Deutsche Forschungsgemeinschaft (DFG) - Project number 100308792
 
Final Report Year 2013

Final Report Abstract

The aim of Z3 was to set up two key genomic technologies for the clinical research unit. The first technology is a high-throughput loss-of-function analysis in mammalian cells; here, cells are infected with pools of lentiviral shRNAs and alterations in abundance of each shRNA are used to measure how each shRNA affects fitness under the specific experimental conditions. Screens are analyzed by high-throughput sequencing. A robust analysis pipeline for these screens has been implemented and has been used to identify shRNAs that affect fitness of multiple myeloma cells in the presence of a MAPkinase inhibitor. The second technology is whole-exome sequencing to identify mutations in the genome of multiple myeloma samples. The technology has been established and 11 exomes have been analyzed. The results point to multiple alterations in receptor tyrosine kinase signaling and in adhesion molecules. The results from both experiments now form the basis for specific scientific projects (TP9N, TP10N) in the next funding period.

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