Naslov
GENERATION OF A NEW MOUSE MUTANT STRAIN WITH INDUCIBLE AND T CELL SPECIFIC CONTROL OF CRE

Autori
Antulov, Ronald ; Mandarić, Sanja ; Zafirova, Biljana ; Polić, Bojan

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
14th International FEBS Summer School on Immunology, IMMUNE SYSTEM: GENES, RECEPTORS AND REGULATION, Hvar, Hrvatska / - , 2007

Skup
14th International FEBS Summer School on Immunology, IMMUNE SYSTEM: GENES, RECEPTORS AND REGULATION

Mjesto i datum
Hvar, Hrvatska, 10.09.2007. - 17.09.2007

Vrsta sudjelovanja
Poster

Vrsta recenzije
Nije recenziran

Ključne riječi
conditional gene targeting; Cre - loxP system; T cells

Sažetak
Conditional gene targeting in mice by inducible Cre/loxP system allows the investigation of the gene function in a cell type specific and inducible manner in vivo. It requires generation of mouse mutants with loxP-flanked gene of interest and inducible expression of Cre recombinase under the control of a cell type specific promoter. To study the role of several genes (i.e.TCRa, NKG2D) in activation and homeostasis of mature T cells in vivo, using tamoxifen inducible CreERT2 system, we have generated a new mouse mutant strain with inducible and T cell specific expression of Cre. T cell specificity and inducibility of Cre were provided by knocking in of the CreERT2 cassette into the CD3e locus. We have designed an appropriate targeting vector that was electroporated into the Bruce4 (C57BL6) embryonic stem (ES) cells. Upon the selection and screening procedure, we have identified several ES cell clones positive for the homologous recombination. Selected ES clones were microinjected into mouse blastocysts and several chimeras were obtained that have given germline transmission of the mutation. To assess the efficiency of tamoxifen inducible Cre mediated deletion in T cells, we crossed CD3-CreERT2 mice to TCRCafl mice, the conditional mouse strain with loxP flanked TCR Ca gene. In this system, we showed TCRa ablation on about 30% of CD4+ and CD8+ T cells mediated by Cre recombination upon tamoxifen administration.

Izvorni jezik
Engleski

Znanstvena područja
Temeljne medicinske znanosti

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