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flk-1-Cre转基因小鼠中内皮细胞特异性Cre重组酶活性。

Endothelium-specific Cre recombinase activity in flk-1-Cre transgenic mice.

作者信息

Licht Alexander H, Raab Sabine, Hofmann Ursula, Breier Georg

机构信息

Department of Molecular Cell Biology, Max Planck Institute for Physiological and Clinical Research, Bad Nauheim, Germany.

出版信息

Dev Dyn. 2004 Feb;229(2):312-8. doi: 10.1002/dvdy.10416.

Abstract

The use of the Cre-loxP recombination system allows the conditional inactivation of genes in mice. The availability of transgenic mice in which the Cre recombinase expression is highly cell type specific is a prerequisite to successfully use this system. We previously have characterized regulatory regions of the mouse flk-1 gene sufficient for endothelial cell-specific expression of the LacZ reporter gene in transgenic mice. These regions were fused to the Cre recombinase gene, and transgenic mouse lines were generated. In the resulting flk-1-Cre transgenic mice, specificity of Cre activity was determined by cross-breeding with the reporter mouse lines Rosa26R or CAG-CAT-LacZ. We examined double-transgenic mice at different stages of embryonic development (E9.5-E16.5) and organs of adult animals by LacZ staining. Strong endothelium-specific staining of most vascular beds was observed in embryos older than E11.5 in one or E13.5 in a second line. In addition, the neovasculature of experimental BFS-1 tumors expressed the transgene. These lines will be valuable for the conditional inactivation of floxed target genes in endothelial cells of the embryonic vascular system.

摘要

Cre-loxP重组系统的应用使得小鼠体内基因能够实现条件性失活。成功使用该系统的一个前提条件是要有Cre重组酶表达具有高度细胞类型特异性的转基因小鼠。我们之前已对小鼠flk-1基因的调控区域进行了表征,这些区域足以使LacZ报告基因在转基因小鼠中实现内皮细胞特异性表达。将这些区域与Cre重组酶基因融合,并构建了转基因小鼠品系。在所得的flk-1-Cre转基因小鼠中,通过与报告基因小鼠品系Rosa26R或CAG-CAT-LacZ杂交来确定Cre活性的特异性。我们通过LacZ染色检查了处于胚胎发育不同阶段(E9.5-E16.5)的双转基因小鼠以及成年动物的器官。在一条品系中,E11.5以上的胚胎以及另一条品系E13.5以上的胚胎中,大多数血管床均观察到强烈的内皮细胞特异性染色。此外,实验性BFS-1肿瘤的新生血管表达了转基因。这些品系对于胚胎血管系统内皮细胞中floxed靶基因的条件性失活将具有重要价值。

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