Expertise

Ihr Ansprechpartner
Dr. Frank Weise
Target-Expressisonssysteme

Tel: 07121 51530-56
weise(at)nmi.de

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 (2001-2015):

>250 cDNA expression vectors generated

>1.200 shRNA vectors for >250 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, Hot Screen GmbH, Novalung GmbH, 4base lab GmbH, NMI TT GmbH

Academia:

KIT (Karlsruhe Institute of Technology), University of Tübingen, University of Stuttgart