Characterization of bacterial artificial chromosome transgenic mice expressing mCherry fluorescent protein substituted for the murine smooth muscle α-actin gene

Authors

  • John J. Armstrong,

    1. Interdepartmental Graduate Program in Cellular and Molecular Biology, Baylor College of Medicine, One Baylor Plaza, Houston, Texas
    2. Center for Cell and Gene Therapy, Baylor College of Medicine, One Baylor Plaza, Houston, Texas
    3. Children's Nutrition Research Center, Baylor College of Medicine, One Baylor Plaza, Houston, Texas
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  • Irina V. Larina,

    1. Department of Molecular Physiology and Biophysics, Baylor College of Medicine, One Baylor Plaza, Houston, Texas
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  • Mary E. Dickinson,

    1. Department of Molecular Physiology and Biophysics, Baylor College of Medicine, One Baylor Plaza, Houston, Texas
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  • Warren E. Zimmer,

    1. Department of Systems Biology and Translational Medicine, Texas A&M University Health Science Center, College Station, Texas
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  • Karen K. Hirschi

    Corresponding author
    1. Interdepartmental Graduate Program in Cellular and Molecular Biology, Baylor College of Medicine, One Baylor Plaza, Houston, Texas
    2. Center for Cell and Gene Therapy, Baylor College of Medicine, One Baylor Plaza, Houston, Texas
    3. Children's Nutrition Research Center, Baylor College of Medicine, One Baylor Plaza, Houston, Texas
    4. Department of Molecular Physiology and Biophysics, Baylor College of Medicine, One Baylor Plaza, Houston, Texas
    5. Department of Pediatrics, Baylor College of Medicine, One Baylor Plaza, Houston, Texas
    6. Department of Molecular and Cellular Biology, Baylor College of Medicine, One Baylor Plaza, Houston, Texas
    • Molecular Physiology and Biophysics, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030, USA
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Abstract

Smooth muscle α actin (SMA) is a cytoskeletal protein expressed by mesenchymal and smooth muscle cell types, including mural cells (vascular smooth muscle cells and pericytes). Using Bacterial Artificial Chromosome (BAC) recombineering technology, we generated transgenic reporter mice that express a membrane localized cherry red fluorescent protein (mCherry), driven by the full-length SMA promoter and intronic sequences. We determined that the founders and F1 progeny of five independent lines contain 1–3 copies of the mCherry-substituted BAC vector. Furthermore, we characterized the expression of SMA-mCherry in relation to endogenous SMA in the embryo and in adult tissues, and found that the transgenic reporter in each line recapitulated endogenous SMA expression at all time points. We were also able to isolate SMA expressing cells from embryonic tissues using fluorescence-activated cell sorting (FACS). We demonstrated that this marker can be combined with other vital fluorescent reporters and it can be used for live imaging of embryonic cardiodynamics. Therefore, these transgenic mice will be useful for isolating live SMA-expressing cells via FACS and for studying the emergence, behavior, and regulation of SMA-expressing cells, including vascular smooth muscle cells and pericytes throughout embryonic and postnatal development. genesis 48:457–463, 2010. © 2010 Wiley-Liss, Inc.

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