Naslov
Generation of mice with inducible T-cell specific expression of Cre

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

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

Izvornik
Book of Abstacts 2008 ANNUAL MEETING OF THE CROATIAN IMMUNOLOGICAL SOCIETY / Sabina Rabatić - Zagreb, 2008, 24-24

Skup
2008 ANNUAL MEETING OF THE CROATIAN IMMUNOLOGICAL SOCIETY

Mjesto i datum
Šibenik, Hrvatska, 09.09.2008. - 12.09.2008

Vrsta sudjelovanja
Poster

Vrsta recenzije
Domaća recenzija

Ključne riječi
inducible; T cell-specific; CD3eCreERT2

Sažetak
Inducible Cre/lox P system represents an effective tool to study the role of specific gene in vivo, providing spatial (cell type specific) and temporal (inducible) control of gene expression at the same time. Here, we report the generation and characterization of a mouse mutant strain with inducible T cell-specific control of Cre, which we have developed to study the role of several genes in the activation and homeostasis of mature T cells in vivo. We used inducible CreER T2 system where the activity of CreER fusion protein can be induced by administration of tamoxifen. T cell specificity and inducibility of Cre were provided by knocking of the CreERT2 cassette into the CD3e locus. We designed an appropriate targeting vector that was electroporated into the Bruce4 (C57BL6) embryonic stem (ES) cells. Upon the selection and screening procedure, we identified several ES cell clones positive for the homologous recombination. Selected ES clones were microinjected into mouse blastocysts and several chimeras were obtained that gave the germline transmission of the mutation. We crossed Cd3e-CreERT2 mice to TCR Cafl mice, containing loxP flanked TCR Ca region, in order to examine the efficiency of tamoxifen inducible Cre mediated deletion in T cells. In this system, we showed conditional TCRab ablation on about 30% of CD4+ and CD8+ T cells mediated by Cre recombination upon tamoxifen administration. The specificity of Cre expression and quantification of Cre mediated deletion in different cell types and tissues were determined by quantitative PCR method.

Izvorni jezik
Engleski

Znanstvena područja
Temeljne medicinske znanosti

POVEZANOST RADA