Using a histone yellow fluorescent protein fusion for tagging and tracking endothelial cells in ES cells and mice

Authors

  • Stuart T. Fraser,

    1. Department of Medicine, Mount Sinai School of Medicine, New York, New York
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    • The first two authors contributed equally to this work.

  • Anna-Katerina Hadjantonakis,

    1. Developmental Biology Program, Memorial Sloan-Kettering Cancer Center, New York, New York
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    • The first two authors contributed equally to this work.

  • Kenneth E. Sahr,

    1. Department of Medicine, Mount Sinai School of Medicine, New York, New York
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  • Stephen Willey,

    1. Department of Medicine, Mount Sinai School of Medicine, New York, New York
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  • Olivia G. Kelly,

    1. Department of Molecular Physiology and Biophysics, Baylor College of Medicine, Houston, Texas
    2. Biological Imaging Center, California Institute of Technology, Pasadena, California
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  • Elizabeth A.V. Jones,

    1. Biological Imaging Center, California Institute of Technology, Pasadena, California
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  • Mary E. Dickinson,

    1. Department of Molecular Physiology and Biophysics, Baylor College of Medicine, Houston, Texas
    2. Biological Imaging Center, California Institute of Technology, Pasadena, California
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  • Margaret H. Baron

    Corresponding author
    1. Department of Medicine, Mount Sinai School of Medicine, New York, New York
    2. Department of Molecular, Cellular and Developmental Biology, Mount Sinai School of Medicine, New York, New York
    3. Department of Oncological Sciences, Mount Sinai School of Medicine, New York, New York
    4. Department of Cell and Gene Medicine, Mount Sinai School of Medicine, New York, New York
    • Mount Sinai School of Medicine, Box 1079, Departments of Medicine and Molecular, Cell & Developmental Biology, 1425 Madison Ave. 11-70B, New York, NY 10029
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Abstract

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 Flk1 promoter and intronic enhancer. The Flk1::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. genesis 42:162–171, 2005. © 2005 Wiley-Liss, Inc.

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