Cre-mediated site-specific recombination in zebrafish embryos

Authors

  • Ryan Thummel,

    Corresponding author
    1. Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, Kansas
    2. Center for Zebrafish Research and Department of Biological Sciences, University of Notre Dame, Notre Dame, Indiana
    • Center for Zebrafish Research and Department of Biological Sciences, University of Notre Dame, Notre Dame, IN 46566
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  • Christopher T. Burket,

    1. Center for Zebrafish Research and Department of Biological Sciences, University of Notre Dame, Notre Dame, Indiana
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  • Jeffrey L. Brewer,

    1. Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, Kansas
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  • Michael P. Sarras Jr,

    1. Department of Anatomy and Cell Biology, University of Kansas Medical Center, Kansas City, Kansas
    Current affiliation:
    1. Department of Cell Biology and Anatomy, Rosalind Franklin University of Medicine and Science, North Chicago, IL
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  • Li Li,

    1. Department of Anatomy and Cell Biology, University of Kansas Medical Center, Kansas City, Kansas
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  • Martin Perry,

    1. Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, Kansas
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  • Jeffrey P. McDermott,

    1. Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, Kansas
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  • Brian Sauer,

    1. Stowers Institute for Medical Research, Kansas City, Missouri and Department of Biochemistry and Molecular Biology, University of Kansas, Medical Center, Kansas City, Kansas
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  • David R. Hyde,

    1. Department of Anatomy and Cell Biology, University of Kansas Medical Center, Kansas City, Kansas
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  • Alan R. Godwin

    1. Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, Kansas
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Abstract

Cre-mediated site-specific recombination has become an invaluable tool for manipulation of the murine genome. The ability to conditionally activate gene expression or to generate chromosomal alterations with this same tool would greatly enhance zebrafish genetics. This study demonstrates that the HSP70 promoter can be used to inducibly control expression of an enhanced green fluorescent protein (EGFP) –Cre fusion protein. The EGFP–Cre fusion protein is capable of promoting recombination between lox sites in injected plasmids or in stably inherited transgenes as early as 2 hr post–heat shock induction. Finally, the levels of Cre expression achieved in a transgenic fish line carrying the HSP70-EGFPcre transgene are compatible with viability and both male and female transgenic fish are fertile subsequent to induction of EGFP–Cre expression. Hence, our data suggests that Cre-mediated recombination is a viable means of manipulating gene expression in zebrafish. Developmental Dynamics 233:1366–1377, 2005. © 2005 Wiley-Liss, Inc.

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