Mouse Genetics Engineering Center  


  HEADDr. Francina LANGA VIVES / francina.langa-vives@pasteur.fr
  MEMBERSAnne CARBON, Gaëlle CHAUVEAU LE-FRIEC, Abokouo ZAGO


  Annual Report

The Mouse Genetics Engineering Center (Centre d'Ingénierie Génétique Murine, CIGM) is the transgenic mouse core facility of Institut Pasteur. Since its creation in 2003, the CIGM is attached to the Developmental Biology Department.

The CIGM aims to generate new models of transgenic mice by classical transgenesis (pronuclear microinjection of DNA fragments, BACs or YACs) 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 2008, 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 2008, we pursued the technique of classical transgenesis mediated by lentivirus. This technique, generally more efficient than pronuclear microinjection, is based on subzonal microinjection of fertilized eggs with viral particles containing small transgenes (<7.5kb). In 2008 we developed this technique to rat transgenesis. Thus, for a neuroscience research project, we successfully microinjected lentiviral transgenes in WISTAR rat embryos, and we obtained the first transgenic rat “founders” generated at CIGM (Fig 1).

The homologous recombination technique in ES cells and the subsequent microinjection of these modified ES cells into mouse blastocysts allow the generation of germ-line chimeras for targeted transgenesis in the endogenous gene. We obtained our first germ-line chimera in June 2004 (Fig. 2) and since then, we have generated 27 heterozygous/homozygous mice lines corresponding to new KO or KI mouse models.

New tools have been developed in 2008 for targeted transgenesis, including generation of new germ-line chimeras from different 129-derived ES cell lines (129Ola, AB1, CK35Rosa ERT2) and also from C57BL/6 ES cells.

Fig 1. First transgenic rats born at CIGM (2008) Fig 2. First germ-line chimera born at CIGM (2004)

Finally, since its creation, CIGM keeps interacting with a steadily increasing list of research groups made up of 29 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 embryos, Transgenesis, Gene Targeting, Homologous Recombination, Embryonic Stem (ES) cells, Microinjection, DNA transgene, BAC, Lentivirus

Cigm.jpg

Fig 1. First transgenic rats born at CIGM (2008)
Fig 2. First germ-line chimera born at CIGM (2004)



  Publications

Disson O, Grayo S, Huillet E, Nikitas G, LangaF, Dussurget O, Ragon M, Le Monnier A, Babinet C, Cossart P, Lecuit M (2008). Conjugated action of two especie-specific invasion proteins for fetoplacental listeriosis. Nature455: 1114-1118

Lochner M, Peduto L, Cherrier M, Sawa S, Langa F, Varona R, Riethmacher D, Si-Tahar M, Di Santo JP, Eberl G (2008). Equilibrium in vivo of pro-inflammatory IL-17+ and regulatory IL-10+ FoxP3+ RORGt+ T cells. J Exp Med 205(6): 1381-1393

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.





Activity Reports 2009 - Institut Pasteur
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