The Mouse Genetics Engineering Center (Centre d’Ingénierie Génétique Murine, CIGM) is the Institut Pasteur’s transgenic mouse core facility. Since its creation in 2003, the CIGM aims to generate new models of transgenic mice by classical or targeted transgenesis. Classical (additive) transgenesis, which includes lentiviral transgenesis, is based on a random insertion of an exogenous gene. Targeted transgenesis consists in a deletion or a specific modification in an endogenous locus using homologous recombination in Embryonic Stem (ES) cells or, more recently, by directly using specific nucleases. Preferentially used by the Institut Pasteur scientific community, the services of CIGM are also available to outside customers interested in new transgenic, knock-down, knock-out (KO) or knock-in (KI) mouse/rat models. The core facility also provides expert advice regarding the design of gene constructs suitable for additive/targeted transgenesis and homologous recombination experiments.
We have an increasing experience in modification on rat genome using lentiviral vectors and Zinc Finger Nucleases (ZFNs). Rat models are more convenient for surgery, due to their medium size, and constitute a robust […]
Genome modifications by CRISPR/cas9 system
The microinjection of the CRISPR/Cas9 system in one-cell embryos is actually revolutionizing the in vivo genome-engineering field because of its big efficiency for KO/ KI (when a donor DNA matrix co-injected with the CRISPR/CAS9 […]
Gene editing by specific nucleases (zinc finger nucleases-zfns, tale nucleases, CRISPR/Cas9 system)
The nucleases (ZFN pairs, TALE pairs, Cas9/CRISPR system) are microinjected, mainly in its mRNA form, into the pronucleus and/or the cytoplasm of one-cell mouse/rat embryos in order to get the desired targeted modification. These […]
Microinjection of “External” Mutant ES Cells And Subsequently Germ-Line Chimera Generation
The easy availability of modified ES cells obtained from KOMP and EUCOMM high-throughput gene-targeting mutagenesis programs (mainly JM8.N4, JM8 A3.N1 ES cell lines) has considerably increased the requests for these direct microinjections in the […]
Lentiviral transgenesis is a special type of classical/additive transgenesis. The transgene, inserted in a lentiviral construct, is microinjected directly under the zona pellucida of one-cell embryos. Then viruses are able to enter directly the […]
Targeted Transgenesis By Homologous Recombination In Embryonic Stem (ES) Cells
A gene targeting vector is transfected into the desired mouse ES cells to modify an endogenous gene by homologous recombination. The mutant ES cells thus obtained are subsequently microinjected into mouse blastocysts for the […]
Classical/Additive Transgenesis by Microinjection
This type of transgenesis consists in the microinjection of the gene of interest into the pronucleus of fertilized eggs to generate transgenic mice expressing the transgene. The microinjected transgenes, which integrated randomly into the […]
Microscope +Transferman NK2 Micromanipulators and Femtojet, Cell Tram Air, Cell Tram Oil MIcroinjectors. The system (manual at the microscope level) allow, like the ASTP station above-mentioned, the storage and recall of positions and the […]
The diascopic stereomicroscope allows embryo counting and the preparation of the reimplantation pipette. The stereomicroscope coupled to a coaxial Episcopic Illuminator (Fibber Optic Light Source) offers bright illumination over the entire surface of the […]
The electroporator (with the appropriated parameters) gives the necessary pulse for the entering of the gene targeting vector or recombinase-expressing plasmid into the Embryonic Stem (ES) Cells.
The laser module, attached just like a typical objective, permits non-contact ablation of targeted membranes. It is mainly used in transgenesis for ablation of the zona pellucida for microinjection of eight-cell and morula stage […]
The piezo-assisted microinjection increases success rates for transgenesis by IntraCytoplasmic Sperm Injection (ICSI), where sperm are coated with exogenous DNA and injected into oocytes. The piezo impact drive, attached to the injecting micropipette holder, […]
Station including Transferman NK2 Micromanipulators and Femtojet, Cell Tram Air, Cell Tram Oil MIcroinjectors. The system combines the operation of both microscope and micromanipulators into one user-oriented control unit (first motorized integrated transgenic platform). […]
2014Human hematopoietic reconstitution and HLA-restricted responses in nonpermissive alymphoid mice, J. Immunol. 2014 Aug;193(3):1504-11.
2014Associations of HLA-A, HLA-B and HLA-C alleles frequency with prevalence of herpes simplex virus infections and diseases across global populations: implication for the development of an universal CD8+ T-cell epitope-based vaccine, Hum. Immunol. 2014 Aug;75(8):715-29.
2014Protective role of LGP2 in influenza virus pathogenesis, J. Infect. Dis. 2014 Jul;210(2):214-23.
2013HLA-A*01:03, HLA-A*24:02, HLA-B*08:01, HLA-B*27:05, HLA-B*35:01, HLA-B*44:02, and HLA-C*07:01 monochain transgenic/H-2 class I null mice: novel versatile preclinical models of human T cell responses, J. Immunol. 2013 Jul;191(2):583-93.
2013Penicillin resistance compromises Nod1-dependent proinflammatory activity and virulence fitness of neisseria meningitidis, Cell Host Microbe 2013 Jun;13(6):735-45.
2012Generation and characterization of a tamoxifen inducible Msx1(CreERT2) knock-in allele, Genesis 2013 Feb;51(2):110-9.
2012The four and a half LIM-only protein 2 regulates liver homeostasis and contributes to carcinogenesis, J. Hepatol. 2012 Nov;57(5):1029-36.
2012Loss of central and peripheral CD8+ T-cell tolerance to HFE in mouse models of human familial hemochromatosis, Eur. J. Immunol. 2012 Apr;42(4):851-62.
2009Characterization of the thymic IL-7 niche in vivo, Proc. Natl. Acad. Sci. U.S.A. 2009 Feb;106(5):1512-7.
2008Conjugated action of two species-specific invasion proteins for fetoplacental listeriosis, Nature 2008 Oct;455(7216):1114-8.
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