Target expression systems
- Syntenin overexpression
The Target Expression Systems group focuses on the functional analysis of proteins within the scope of target identification / target validation for the pharmaceutical industry. To this end, we apply a panel of gene transfer methods in order to overexpress or knock-down gene expression in eukaryotic cells.
Furthermore, these techniques are used to unravel the mode of action of new compounds.
Track record: Expertise in gene transfer and expression analysis (2001-2010):
>250 cDNA expression vectors generated
>800 shRNA vectors for 155 targets validated
Techniques, methods, equipment
RNA interference (RNAi)
- siRNA
- shRNA
- miRNA
Gene transfer
- plasmid-based
- adeno-, retro-, lentiviral (biosafety level 2 laboratory)
- nucleofection (amaxa®)
Gene expression
- qRT-PCR (TaqMan™)
Protein expression analysis
- Western blot
- immunoprecipitation
- immunofluorescence microscopy
Protein localisation studies
- confocal microscopy
- immunohistochemistry
Analysis of protein function
- Determination of the induction of apoptosis
- Determination of the cell viability
- Signal transduction analysis
- Various reporter assays
Fee-for-service packages
Gene overexpression/knock-down:
- Gene cloning (isolation of mRNA, cDNA synthesis, amplification and cloning of open reading frames)
- Generation and application of viral and non-viral vectors to overexpress or knock-down genes in eukaryotic cell cultures as well as in primary cell cultures
- Optimisation of viral transduction
- Generation of stably expressing cell lines (polyclonal and/or monoclonal) in conjunction with expression analysis at the mRNA level and at the protein level, followed by functional characterisation
- Development of improved, more physiological cell cultur models (co-cultures, cell stress/injury paradigms)
- Overexpression and subsequent purification of proteins in laboratory scale (ca. 1 mg of protein) using bacterial and eukaryotic cell systems
Network
Industry:
Bayer Healthcare Pharmaceuticals, EDI GmbH, Novalung GmbH, 4base lab GmbH, NMI TT GmbH
Academia:
KIT (Karlsruhe Institute of Technology), University of Tübingen, University of Stuttgart