Practical HP-F4: Advanced Methods in Cell Biology
Type: Practical with Student Seminar
Date: 30.01.-17.02.2012
Hosts/Supervisors: Frank Lyko, Achim Breiling, Harald Herrmann, Michael Boutros, Dominique Kranz (contact: h.herrmann@dkfz.de)
Mitotic human U2OS osteosarcoma cell stained for A-type lamins revealing the complete segregation of lamin A and lamin C
| © dkfz.de Topics:
- A) Analyzing and modulating DNA methylation patterns in human cancer cells
- B) Investigation of protein topogenesis in cultured cells by immunofluorescence microscopy, differential cell extraction and cDNA transfection
- C) RNA interference as a tool to study gene function
Content:
General: cell culture, immunofluorescence microscopy, Western blotting, cDNA transfection, bisulfite sequencing, RNAi screening
Details for part C): RNA interference (RNAi) has become an important method for the characterization of gene function. RNAi allows the silencing of specific genes through introduction of short double-stranded RNAs homologous to endogenous mRNAs. As part of the course, participants will learn how to perform RNAi experiments in human cells and analyse phenotypic changes using different cell-based assays. We will perform a small scale RNAi screen to analyze the Wnt signaling pathways which are known to be involved in the development of various cancers.
