Supervisor : Prof. Dr. Claude LIBERT
Tel: +32 9 33 13 639/744 - Fax:+32 9 221 76 73
E-mail: Tino.Hochepied@irc.vib-UGent.be Leen.Vanhoutte@irc.vib-UGent.be
The Transgenic Mice Core Facility (TMCF) of the IRC was established in 2004 to cater to the needs of IRC investigators. The mission of the TMCF is to provide efficient and economic access to transgenic animals and related technologies.
Summary of services
- Gene targeting in ES-cells
Embryonic Stem (ES) cells are electroporated with a targeting vector designed to introduce a specific mutation in the mouse genome. After antibiotic selection, surviving colonies are picked and expanded for screening and freezing. To generate transgenic mice over-expressing a gene of interest, dedicated ES-cells and targeting vectors are available to specifically introduce the transgene in the Rosa26 locus.
- Generation of chimeras
Correctly targeted ES cells are introduced in an early mouse embryo. This can be done by injecting the ES cells into blastocysts or by aggregating ES cell clumps with morulas; The ES cells assist in the formation of the fetus. The resulting mouse is a chimera composed of two cell types: cells derived from the embryo and those derived from the introduced ES cells. The chimeric mice are mated with wild-type mice to transmit the desired mutation from the chimeric to the offspring.
- Generation of mutant mice
Nuclease mediated gene editing is a powerful way for making mutant mice. Crispr/Cas9 technology is very efficient for making knockin and even multiplex knockout mice. The technology also allows for a significant time reduction compared to ES-cell mediated gene targeting and can be used to generate mutant mice in virtually any desired background.
- Rederivation of mouse strains
The presence of any type of mouse pathogen (whether it causes clinical disease or not), changes the physiology of the mouse, which can lead to false experimental results. Mouse strains harboring pathogens can be ‘rederived’ to a pathogen-free status by transferring pre-implantation embryos from a contaminated mouse to a pathogen-free foster mother. The resulting pups will then also be pathogen-free.
The purpose of cryopreservation is to protect against loss of valuable, unique mouse lines through breeding failure or disease, and to eliminate the cost of maintaining mouse lines that are not in use. Sperm freezing is the default method as it is quick and few mice are needed. An IVF is required with the frozen sperm to revive the mouse line. For complex genetic backgrounds, embryo freezing is the preferred method.
- Haenebalcke, et al. The ROSA26-iPSC mouse: A conditional, inducible and exchangeable resource for studying cellular (de)differentiation.
Cell Rep. 3, 335-341, 2013.
- Vanhooren, et al. Mice overexpressing beta-1,4-galactosyltransferase I are resistant to TNF-induced inflammation and DSS-induced colitis.
PLoS One. 8, e79883, 2013.
- Vyas, et al. RNF4 is required for DNA double-strand break repair in vivo.
Cell Death Differ. 20, 490-502, 2013.
- Denecker et al. Caspase-14 protects against epidermal UVB photo-damage and water loss.
Nature Cell Biol 9, 666-674, 2007.
- Hochepied et al. Breaking the species barrier: Derivation of germline-competent embryonic stem cells from (Mus spretus × C57BL/6) hybrids.
Stem Cells 22, 441-447, 2004.