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Generation of genetically modified mice by random insertion of a transgene can be performed by pronuclear DNA microinjection. This technique is a cost-effective and can be fast alternative for generation of knock down, transgene over expression and reporter lines.

A solution containing the transgene is injected directly into the male pronucleus of fertilized oocystes. The eggs are implanted in the oviduct of pseudopregnant recipient mice. At birth the newborn are tested for the transgene detection. All positive animals (called the founders) will be different in their number of copies and transgenes insertion sites.

ICS expertise:

  • Founders will be analyzed by qPCR to determine number of transgenic copies.
  • Molecular characterization of the lines generated, including Southern Blot, qRT-PCR or Western Blot, can be done upon request by our gene expression service.
  • C57BL/6N, C57BL/6J and FVB/N background are used in routine. Other backgrounds may be available upon request.

Risks:

  • Integration of the transgene is random. Disturbing endogenous gene(s) expression is possible.
  • The expression pattern will need to be evaluated for each founder as it may differ depending of the site(s) of integration of the transgene.

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