期刊
GENESIS
卷 42, 期 3, 页码 162-171出版社
WILEY
DOI: 10.1002/gene.20139
关键词
Flk1 :: H2B-EYFP; endothelial cells; fluorescent protein; EYFP; vasculogenesis; angiogenesis; imaging; transgenic mice; histone fusion
资金
- NHLBI NIH HHS [R01 HL077187, R01 HL062248] Funding Source: Medline
- NIBIB NIH HHS [R01 EB002209] Funding Source: Medline
- NIDDK NIH HHS [R01 DK052191] Funding Source: Medline
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.
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