Production of protein-tagged pluripotent und differentiated ES cells

Coordinator:		Prof. Dr. Wolfgang Wurst
Institution:		Institut für Entwicklungsgenetik, Helmholtz Zentrum München
Homepage:		www.helmholtz-muenchen.de

This subproject will perform the targeting of the ES cells by electroporation of the tagging constructs, it will identify the targeted clones, and it will also store the clones and distribute them to the other subprojects of DiGtoP.
Mouse ES cell lines harboring tagged proteins will be differentiated into different neuronal, cardiac and pancreatic cells. To this end, we will implement standard differentiation protocols for the different lineages. To facilitate the production of homogeneous lineage-specific ES cell cultures we will establish mES cell lines stably carrying lineage-specific selection markers, if necessary. This will allow the selection of the desired cell type directly by selection or by fluorescence-activated cell sorting (FACS).
The tagged protein interaction partners will be isolated from the corresponding cell population and determined by mass spectrometry in SP6.
In parallel, human ES cell lines will be tagged and differentiated into neural, cardiac and pancreatic cell lineages in SP5 allowing to compare the human and mouse cell type-specific interactions.




Additional relevant Internet links:
DiGTOP