Kurzfassung

This laboratory is focused on signal transduction and regulation of gene expression in both human and murine primary T cells and murine mast cells.
It covers all aspects of molecular cloning, primer design, qRT-PCR, PCR-based genotyping of transgenic mice, expression of recombinant proteins and. In particular, techniques have been established which allow the functional analyses of proteins which is a conditio sine qua non for the evaluation of target mRNAs and proteins identified by genomic...This laboratory is focused on signal transduction and regulation of gene expression in both human and murine primary T cells and murine mast cells.
It covers all aspects of molecular cloning, primer design, qRT-PCR, PCR-based genotyping of transgenic mice, expression of recombinant proteins and. In particular, techniques have been established which allow the functional analyses of proteins which is a conditio sine qua non for the evaluation of target mRNAs and proteins identified by genomic and proteomic approaches. This includes design, and functional testing of siRNA molecules for “knock down” experiments. Biological activity of individual siRNA molecules can be assayed by qRT-PCR, FACS analyses and western blotting (if antibody is available).
In order to study the activation and regulation of specific promoters and transcription factors, chromatin immunoprecipitation (ChIP), reporter gene assays and overexpression experiments can also be performed. The latter manipulations are based on procedures, optimized in this laboratory, which allow high efficiency non-viral transfections of primary human and, for the first time, conventional CD4+, CD8+ and regulatory murine T cells and mast cells with siRNA molecules and plasmids.
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