Gene Targeting Unit

The Stem Cell Center – Gene Targeting Unit has been set up for handling and manipulating mouse embryonic stem (ES) cells. By combining homologous recombination, site-specific recombination and transgenesis, genetic changes described in human disease can be copied into the mouse genome, thus engineering ideal disease models.

ES cell gene targeting
The several missions of the ES cell core facility include the production of quality-controlled ES cell lines with mutations introduced by homologous recombination using a targeting vector provided by the researcher, the creation and handling of quality-controlled ES cell lines using state-of-the-art reagents, and enhancing knowledge of mouse genetics, ES cell culture and manipulation. Procedures for the manipulation of ES cells and the generation of mice are labour-intensive and technically demanding. Facilitating experiments of this type is one of the purposes of the Unit. Targeted ES cells can be used to generate germline ES cell-mouse chimeras that can be bred to generate mouse lines or for in vitro assays. We also develop in vivo inducible gene targeting systems focusing on breast cancer using female ES cell lines, and try to generate new transgenic recombinase mouse models.

Gene targeting tool box
We maintain a “gene targeting tool box” in addition to providing expertise, cell lines, efficient exchange of information, and stem cell assays. The collection includes Cre and Flp recombinase-expressing transgenic lines and Cre and Flp activity-reporter lines, which are essential for the generation and characterization of conditional, inducible, and/or tissue-specific mutant mice. The Unit also maintains a collection of “ES and vectors tools”, reagents, feeders (neomycin resistant or neo/hygro/puro resistant), cell lines, and plasmid vectors.

Derivation of novel ES cell lines
The availability of highly germline-competent ES cells derived from inbred strains, which are preferentially used in biomedical research, would greatly facilitate the generation of targeted mutations in appropriate genetic backgrounds. We now have the expertise and skills to isolate and culture inbred ES cells, such as FVB/N and C57BL6/N ES cell lines. This provides essential tools for analysis of the mutations, especially when the phenotype is embryonic lethal at the early stages of development. Currently our interest is focused on generating induced pluripotent stem cells.

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