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Use of gene-specific nucleases for the generation of genetically engineered rats for the analysis of immune responses

Mardi 27 février 2018 11:00 - Duree : 1 heure
Lieu : Salle de Conférence de l’IAB - Rond Point de La Chantourne, 38700 La Tronche (arrêt de tram Grand Sablon, ligne B)

Orateur : Ignacio ANEGON (Université de Nantes)

The rat is widely used as an experimental model and despite the availability of many transgenic rats the lack of KO and KI mutants has been an obstacle to fully develop the potential of rat models. Since 2009, ZFNs have been used to readily generate several KO rats, among them for heavy and light chain antibody genes. These rats were then crossed with transgenic rats in which human immunoglobulin genes were introduced by BAC and YAC microinjection. These immunoglobulin humanized rats showed human V gene diversity comparable to the one of human antibodies and monoclonal antibodies of high affinity were generated from these rats using hybridoma technology. We then used these rats to generate monoclonal antibodies using a new technique based on single B cell antigen-specific sorting followed by expression cloning of the immunoglobulin genes of these B cells. I-CreI-engineered meganucleases were used to generate rats KO for Rag1 with a SCID phenotype. TALENs were used to generate rats KO for the gamma chain of the IL2 receptor (Il2rg) (Ménoret et al. manuscript in preparation). Rag1 and Il2rg KO rats were crossed to obtain double KO rats (RRG) with a more pronounced T, B and NK immunodeficient phenotype that each single KO. To inhibit phagocytosis of human cells by rat macrophages and thus facilitate immune system humanization, we generated human SIRPalpha transgenic rats using piggyBac mediated BAC transgenesis that are being crossed with RRG animals. We also used TALENs to generate dystrophin KO rats to reproduce Düchenne disease and we have characterized the immunophenotype of the muscle of these animals (Ouisse et al. submitted). The availability of gene-specific nucleases provide an extremely useful tool to generate KO and KI rats for the analysis of gene function, generate disease models and biotechnological applications in the area of immunology.

Contact : karin.sadoul@univ-grenoble-alpes.fr

Discipline évènement : (Biologie / Chimie)
Entité organisatrice : (IAB)
Nature évènement : (Séminaire)
Evènement répétitif : (Mardis de l’IAB)
Site de l'évènement : Pôle Santé / La Tronche

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