Tie-1-directed expression of Cre recombinase in endothelial cells of embryoid bodies and transgenic mice.
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Tie-1-directed expression of Cre recombinase in endothelial cells of embryoid bodies and transgenic mice. / Gustafsson, E; Brakebusch, C; Hietanen, K; Fässler, R.
I: Journal of Cell Science, Bind 114, Nr. Pt 4, 2001, s. 671-6.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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TY - JOUR
T1 - Tie-1-directed expression of Cre recombinase in endothelial cells of embryoid bodies and transgenic mice.
AU - Gustafsson, E
AU - Brakebusch, C
AU - Hietanen, K
AU - Fässler, R
N1 - Keywords: Animals; Embryo, Mammalian; Endothelium; Gene Expression Regulation, Enzymologic; Integrases; Mice; Mice, Transgenic; Receptor Protein-Tyrosine Kinases; Receptor, TIE-1; Receptors, Cell Surface; Receptors, TIE; Recombination, Genetic; Viral Proteins
PY - 2001
Y1 - 2001
N2 - Tissue-specific gene inactivation using the Cre-loxP system has become an important tool to unravel functions of genes when the conventional null mutation is lethal. We report here the generation of a transgenic mouse line expressing Cre recombinase in endothelial cells. In order to avoid the production and screening of multiple transgenic lines we used embryonic stem cell and embryoid body technology to identify recombinant embryonic stem cell clones with high, endothelial-specific Cre activity. One embryonic stem cell clone that showed high Cre activity in endothelial cells was used to generate germline chimeras. The in vivo efficiency and specificity of the transgenic Cre was analysed by intercrossing the tie-1-Cre line with the ROSA26R reporter mice. At initial stages of vascular formation (E8-9), LacZ staining was detected in almost all cells of the forming vasculature. Between E10 and birth, LacZ activity was detected in most endothelial cells within the embryo and of extra-embryonic tissues such as yolk sac and chorioallantoic placenta. Ectopic expression of Cre was observed in approximately 12-20% of the adult erythroid, myeloid and lymphoid cells and in subregions of the adult brain. These results show that the tie-1-Cre transgenic strain can efficiently direct deletion of floxed genes in endothelial cells in vivo.
AB - Tissue-specific gene inactivation using the Cre-loxP system has become an important tool to unravel functions of genes when the conventional null mutation is lethal. We report here the generation of a transgenic mouse line expressing Cre recombinase in endothelial cells. In order to avoid the production and screening of multiple transgenic lines we used embryonic stem cell and embryoid body technology to identify recombinant embryonic stem cell clones with high, endothelial-specific Cre activity. One embryonic stem cell clone that showed high Cre activity in endothelial cells was used to generate germline chimeras. The in vivo efficiency and specificity of the transgenic Cre was analysed by intercrossing the tie-1-Cre line with the ROSA26R reporter mice. At initial stages of vascular formation (E8-9), LacZ staining was detected in almost all cells of the forming vasculature. Between E10 and birth, LacZ activity was detected in most endothelial cells within the embryo and of extra-embryonic tissues such as yolk sac and chorioallantoic placenta. Ectopic expression of Cre was observed in approximately 12-20% of the adult erythroid, myeloid and lymphoid cells and in subregions of the adult brain. These results show that the tie-1-Cre transgenic strain can efficiently direct deletion of floxed genes in endothelial cells in vivo.
M3 - Journal article
C2 - 11171372
VL - 114
SP - 671
EP - 676
JO - Journal of Cell Science
JF - Journal of Cell Science
SN - 0021-9533
IS - Pt 4
ER -
ID: 5141765