In situ labelling of disease proteins in embryonic stem cells with gene trap induced multipurpose alleles

Coordinator:		Prof. Dr. Harald von Melchner
Institution:		Molekulare Hämatologie, Goethe Universität Frankfurt
Homepage:		www.vonmelchner.de

This subproject will develop technology for the high throughput insertion of N-terminal protein tags into genes trapped in mouse embryonic stem cells (ESCs) by conditional gene trap vectors. These vectors insert site specific recombinase target sequences along with the gene trapping cassettes into the genome and thereby create multipurpose alleles amenable to postinsertional modifications by recombinase mediated cassette exchange (RMCE). ESC lines with suitable gene trap insertions in the disease genes will be recovered from the existing gene trap resources and subjected to RMCE to insert an appropriate protein tag into the gene trap locus. Tagged proteins expressed from endogenous promoters under various physiological conditions will be subjected to live cell imaging and proteomic interactor mapping by mass spectrometry.








Additional relevant Internet links:
DiGTOP