|Mouse Genetics Engineering Center|
|HEAD||Dr. Francina LANGA VIVES / email@example.com|
|MEMBERS||Anne Carbon / Gaëlle CHAUVEAU / Abokouo ZAGO
Ancient members: Rémi BEAU (until July 2007) / Françoise THOURON (until June 2007)
Created in 2003, the Centre d’Ingénierie Génétique Murine(CIGM)is a technological core facility, which aims at generating new models of genetically-engineered mice either by classical transgenesis or by homologous recombination in embryonic stem (ES) cells (targeted transgenesis).Preferentially open to the Institut Pasteur scientific community, the services of CIGM also are available to outside customers interested in new transgenic, knock-out (KO) or knock-in(KI) mouse models. The core facility also provides expert advice regarding the design of gene constructs suitable for transgenesis and homologous recombination experiments.
The classical transgenesis service involves the microinjection of the gene of interest into the pronuclei of fertilized eggs in order to generate transgenic mice expressing the transgene. The microinjected transgenes include DNA linear fragments (2kb to 20kb) as well as larger DNAs from bacterial or yeast artificial chromosomes (BACs, YACs). In 2007, we have successfully microinjected an increasing number of BACs. Moreover, we increased the use of pure genetic backgrounds as egg-donor mouse strains, including FVB/N, BALB/c and particularly C57BL/6, which is par excellence, the model of choice for studies in neuroscience and immunology. Other mouse strains currently used at CIGM are hybrids, such as B6SJLF1, B6D2F1, B6CBAF1 or B6BALB/cF1.
In addition, we pursued the technique of lentiviral transgenesisby subzonal injection of fertilized eggs with viral particles containing small transgenes (<7.5kb), more efficient than pronuclear injection.
The homologous recombination technique in ES cellsand the subsequent microinjection of these modified ES cells into mouse blastocysts allow the generation of germ-line chimeras for targeted transgenesisin the endogenous gene. We obtained our first germ-line chimera (Fig 1) in June 2004. Since, we have generated 25 heterozygous/homozygous mice lines corresponding to new KOor KImouse models.
Different 129-derived ES cell lines (i.e. CK35, AB1 and TC-1) were used in 2007 for microinjection and generation of mouse chimeras. C57BL/6-derived ES cell lines will be used in 2008.
The CIGM has at its disposal an independent animal room fully equipped with ventilated racks, ensuring a rigorous compliance with the high health standard expected of the new transgenic mice generated. The space in the animal facility has increased by 15% in 2007.
Finally, since its creation, CIGM keeps interacting with a steadily increasing list of research groups made up of 26 Units within the Institut Pasteur (belonging to 7 out of the 10 Departments), and other Research Institutions in France (CNRS Units UMR 7622-Paris 6 and UMR146-Institut Curie-Orsay, INSERM Units E-365-Lariboisière, U787-Pitié-Salpétrière, U-813-Necker and E9935-Robert Debré-Paris, and U522-Pontchaillou-Rennes, the Institut National de la Transfusion Sanguine-Paris) as well as in foreign countries (Université Libre de Bruxelles- Belgium, Universitat de Barcelona-Spain).
Keywords: Mouse, Transgenesis, Homologous recombination, Embryonic Stem (ES) cells, Microinjection
Monet M, Domenga V, Lemaire B, Souilhol C, Langa F, Babinet C, Gridley T, Tournier-Lasserve E, Cohen-Tannoudji M, Joutel A (2007). The archetypal R90C CADASIL-NOTCH3 mutation retains NOTCH3 function in vivo.Hum. Mol. Genet. 16(8): 982-992.
Hyenne V, Souilhol C, Cohen-Tannoudji M, Cereghini S, Petit C, Langa F, Maro B, Simmler MC (2007). Conditional knock-out reveals that zygotic vezatin-null mouse embryos die at implantation.Mech. Dev. 124(6): 449-462.
Delmaghani S, del Castillo FJ, Michel V, Leibovici M, Aghaie A, Ron U, Van Laer L, Ben-Tal N, Van Camp G, Weil D, Langa F, Lathrop M, Avan P, Petit C (2006). Mutations in the gene encoding pejvakin, a novel protein expressed in the afferent auditory pathway, cause DFNB59 auditory neuropathy in man and mouse.Nat Genet 38(7): 770-778.
Langa F (2005). The new core facility «Centre d’Ingénierie Génétique Murine» at Pasteur Institute: Start-up and first results.Transgenic Res. 14: 494.
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Activity Reports 2007 - Institut Pasteur
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