Using a histone yellow fluorescent protein fusion for tagging and tracking endothelial cells in ES cells and mice
Using a histone yellow fluorescent protein fusion for tagging and tracking endothelial cells in ES cells and mice
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DOI：
10.1002/gene.20139
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发表时间：
2005-07-01
期刊：
影响因子：
1.5
通讯作者：
Baron, MH
中科院分区：
文献类型：
--
作者：
Fraser, ST;Hadjantonakis, AK;Baron, MH
我们报道了第一个内皮细胞系特异性转基因小鼠，可以在亚细胞分辨率下进行活体成像。我们制备了一种可用于核小体荧光标记的H2B-EYFP融合蛋白，并将其用于特异性标记小鼠内皮细胞和分化胚胎干细胞(ES)。在FIk1启动子和内含子增强子的控制下，表达了编码组蛋白H_2B的融合基因，并在其C端标记了增强型黄色荧光蛋白(EYFP)。FIk1：：H2B-EYFP转基因小鼠是存活的，并且在发育中的胚胎和成年动物的内皮细胞中保持着高水平的染色质定位报告表达。在分化的ES细胞和胚胎中，荧光的开始与内皮细胞特化的开始相对应。这些转基因品系允许对正常和病理性血管生成和血管生成进行实时成像，以跟踪单个细胞和有丝分裂事件，其详细程度在小鼠中是前所未有的。
We report the first endothelial lineage-specific transgenic mouse allowing live imaging at subcellular resolution. We generated an H2B-EYFP fusion protein which can be used for fluorescent labeling of nucleosomes and used it to specifically label endothelial cells in mice and in differentiating embryonic stem (ES) cells. A fusion cDNA encoding a human histone H2B tagged at its C-terminus with enhanced yellow fluorescent protein (EYFP) was expressed under the control of an FIk1 promoter and intronic enhancer. The FIk1::H2B-EYFP transgenic mice are viable and high levels of chromatin-localized reporter expression are maintained in endothelial cells of developing embryos and in adult animals upon breeding. The onset of fluorescence in differentiating ES cells and in embryos corresponds with the beginning of endothelial cell specification. These transgenic lines permit real-time imaging in normal and pathological vasculogenesis and angiogenesis to track individual cells and mitotic events at a level of detail that is unprecedented in the mouse.