Site-specific transgenesis in Xenopus

Authors

  • Michael E. Zuber,

    Corresponding author
    1. Department of Ophthalmology, The Center for Vision Research and the SUNY Eye Institute, Upstate Medical University, Syracuse, New York
    2. Department of Neuroscience & Physiology, The Center for Vision Research and the SUNY Eye Institute, Upstate Medical University, Syracuse, New York
    3. Department of Biochemistry & Molecular Biology, The Center for Vision Research and the SUNY Eye Institute, Upstate Medical University, Syracuse, New York
    • SUNY Upstate Medical University, 750 East Adams Street, Syracuse, NY 13210, USA
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  • Heather S. Nihart,

    1. Department of Ophthalmology, The Center for Vision Research and the SUNY Eye Institute, Upstate Medical University, Syracuse, New York
    2. Department of Biochemistry & Molecular Biology, The Center for Vision Research and the SUNY Eye Institute, Upstate Medical University, Syracuse, New York
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  • Xinming Zhuo,

    1. Department of Ophthalmology, The Center for Vision Research and the SUNY Eye Institute, Upstate Medical University, Syracuse, New York
    2. Department of Biochemistry & Molecular Biology, The Center for Vision Research and the SUNY Eye Institute, Upstate Medical University, Syracuse, New York
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  • Sudha Babu,

    1. Department of Ophthalmology, The Center for Vision Research and the SUNY Eye Institute, Upstate Medical University, Syracuse, New York
    2. Department of Biochemistry & Molecular Biology, The Center for Vision Research and the SUNY Eye Institute, Upstate Medical University, Syracuse, New York
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  • Barry E. Knox

    1. Department of Ophthalmology, The Center for Vision Research and the SUNY Eye Institute, Upstate Medical University, Syracuse, New York
    2. Department of Neuroscience & Physiology, The Center for Vision Research and the SUNY Eye Institute, Upstate Medical University, Syracuse, New York
    3. Department of Biochemistry & Molecular Biology, The Center for Vision Research and the SUNY Eye Institute, Upstate Medical University, Syracuse, New York
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Abstract

Transgenesis is an essential, powerful tool for investigating gene function and the activities of enhancers, promoters, and transcription factors in the chromatin environment. In Xenopus, current methods generate germ-line transgenics by random insertion, often resulting in mosaicism, position-dependent variations in expression, and lab-to-lab differences in efficiency. We have developed and tested a Xenopus FLP-FRT recombinase-mediated transgenesis (X-FRMT) method. We demonstrate transgenesis of Xenopus laevis by FLP-catalyzed recombination of donor plasmid cassettes into F1 tadpoles with host cassette transgenes. X-FRMT provides a new method for generating transgenic Xenopus. Once Xenopus lines harboring single host cassettes are generated, X-FRMT should allow for the targeting of transgenes to well-characterized integration site(s), requiring no more special reagents or training than that already common to most Xenopus labs. genesis 50:325–332, 2012. © 2012 Wiley Periodicals, Inc.

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