A facile method for somatic, lifelong manipulation of multiple genes in the mouse liver

Authors

  • Kirk J. Wangensteen,

    1. The Arnold and Mabel Beckman Center for Transposon Research, Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN
    2. Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, MN
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  • Andrew Wilber,

    1. The Arnold and Mabel Beckman Center for Transposon Research, Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN
    2. Gene Therapy Program, Institute of Human Genetics, University of Minnesota, Minneapolis, MN
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  • Vincent W. Keng,

    1. The Arnold and Mabel Beckman Center for Transposon Research, Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN
    2. Cancer Center, University of Minnesota, Minneapolis, MN
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  • Zhiying He,

    1. Department of Laboratory Medicine and Pathology, University of Minnesota, Minneapolis, MN
    2. Laboratory of Molecular Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai Institute for Biological Sciences, Chinese Academy of Sciences, Shanghai, China
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  • Ilze Matise,

    1. College of Veterinary Medicine, University of Minnesota, Minneapolis, MN
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  • Laura Wangensteen,

    1. Department of Medicine, University of Minnesota, Minneapolis, MN
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  • Corey M. Carson,

    1. The Arnold and Mabel Beckman Center for Transposon Research, Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN
    2. Cancer Center, University of Minnesota, Minneapolis, MN
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  • Yixin Chen,

    1. Department of Laboratory Medicine and Pathology, University of Minnesota, Minneapolis, MN
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  • Clifford J. Steer,

    1. Department of Medicine, University of Minnesota, Minneapolis, MN
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  • R. Scott McIvor,

    1. The Arnold and Mabel Beckman Center for Transposon Research, Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN
    2. Gene Therapy Program, Institute of Human Genetics, University of Minnesota, Minneapolis, MN
    3. Cancer Center, University of Minnesota, Minneapolis, MN
    4. Department of Laboratory Medicine and Pathology, University of Minnesota, Minneapolis, MN
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  • David A. Largaespada,

    1. The Arnold and Mabel Beckman Center for Transposon Research, Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN
    2. Cancer Center, University of Minnesota, Minneapolis, MN
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  • Xin Wang,

    1. Department of Laboratory Medicine and Pathology, University of Minnesota, Minneapolis, MN
    2. Stem Cell Institute, University of Minnesota, Minneapolis, MN
    3. Laboratory of Molecular Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai Institute for Biological Sciences, Chinese Academy of Sciences, Shanghai, China
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  • Stephen C. Ekker

    Corresponding author
    1. The Arnold and Mabel Beckman Center for Transposon Research, Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN
    2. Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, MN
    Current affiliation:
    1. Mayo Clinic College of Medicine, Department of Biochemistry/Molecular Biology, 200 1st Street SW, Guggenheim 1321A, Rochester, MN 55905
    • Mayo Clinic College of Medicine, Department of Biochemistry/Molecular Biology, 200 1st Street SW, Guggenheim 1321A, Rochester, MN 55905
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    • fax: 507-284-1767.


  • Potential conflict of interest: Nothing to report.

Abstract

Current techniques for the alteration of gene expression in the liver have a number of limitations, including the lack of stable somatic gene transfer and the technical challenges of germline transgenesis. Rapid and stable genetic engineering of the liver would allow systematic, in vivo testing of contributions by many genes to disease. After fumaryl acetoacetate hydrolase (Fah) gene transfer to hepatocytes, selective repopulation of the liver occurs in FAH-deficient mice. This genetic correction is readily mediated with transposons. Using this approach, we show that genes with biological utility can be linked to a selectable Fah transposon cassette. First, net conversion of Fah−/− liver tissue to transgenic tissue, and its outgrowth, was monitored by bioluminescence in vivo from a luciferase gene linked to the FAH gene. Second, coexpressed short hairpin RNAs (shRNAs) stably reduced target gene expression, indicating the potential for loss-of-function assays. Third, a mutant allele of human α1-antitrypsin (hAAT) was linked to Fah and resulted in protein inclusions within hepatocytes, which are the histopathological hallmark of hAAT deficiency disorder. Finally, oncogenes linked to Fah resulted in transformation of transduced hepatocytes. Conclusion: Coexpression with FAH is an effective technique for lifelong expression of transgenes in adult hepatocytes with applicability to a wide variety of genetic studies in the liver. (HEPATOLOGY 2008;47:1714–1724.)

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