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Pig transgenesis by Sleeping Beauty DNA transposition

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Abstract

Modelling of human disease in genetically engineered pigs provides unique possibilities in biomedical research and in studies of disease intervention. Establishment of methodologies that allow efficient gene insertion by non-viral gene carriers is an important step towards development of new disease models. In this report, we present transgenic pigs created by Sleeping Beauty DNA transposition in primary porcine fibroblasts in combination with somatic cell nuclear transfer by handmade cloning. Göttingen minipigs expressing green fluorescent protein are produced by transgenesis with DNA transposon vectors carrying the transgene driven by the human ubiquitin C promoter. These animals carry multiple copies (from 8 to 13) of the transgene and show systemic transgene expression. Transgene-expressing pigs carry both transposase-catalyzed insertions and at least one copy of randomly inserted plasmid DNA. Our findings illustrate critical issues related to DNA transposon-directed transgenesis, including coincidental plasmid insertion and relatively low Sleeping Beauty transposition activity in porcine fibroblasts, but also provide a platform for future development of porcine disease models using the Sleeping Beauty gene insertion technology.

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Acknowledgments

We thank Peter Angels for providing pUbiC-Jun. We thank Tina Fuglsang and Marianne Gregers Johansen for excellent technical assistance. The work by J.E.J. was carried out in the laboratory of A.L.J. This work was made possible through support from the Danish National Advanced Technology Foundation, the Lundbeck Foundation (grant no. R19-A2108), the Danish Research Agency (grant no. 274-05-0197), the Danish Medical Research Council, the Novo Nordisk Foundation, the Carlsberg Foundation, the Danish Cancer Society, the Danish Heart Association, Aage Bangs Foundation, the Augustinus Foundation, and the EU (EU-FP6-STREP, contract number 018961). B.M was funded by a grant from the Danish Cancer Society.

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Authors and Affiliations

  1. Department of Human Genetics, University of Aarhus, Wilh. Meyers Allé 1240, 8000, Aarhus C, Denmark

    Jannik E. Jakobsen, Peter M. Kragh, Brian Moldt, Lars Bolund, Arne Lund Jørgensen, Anders Lade Nielsen & Jacob Giehm Mikkelsen

  2. Department of Genetics and Biotechnology, Faculty of Agricultural Sciences, University of Aarhus, 8830, Tjele, Denmark

    Juan Li, Peter M. Kragh, Lin Lin, Ying Liu, Gábor Vajta & Henrik Callesen

  3. Department of Veterinary Reproduction and Obstetrics, Faculty of Life Sciences, University of Copenhagen, 1870, Frederiksberg C, Denmark

    Mette Schmidt

  4. Danish Agriculture and Food Council, Knowledge Center on Pig Production, 1609, Copenhagen V, Denmark

    Kjeld Dahl Winther

  5. National Veterinary Institute, Technical University of Denmark, 8200, Aarhus N, Denmark

    Brian Dall Schyth

  6. Department of Pathology, Randers Hospital, 8900, Randers, Denmark

    Ida E. Holm

  7. Danish Neuroscience Center, Aarhus University Hospital, 8000, Aarhus C, Denmark

    Ida E. Holm

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  1. Jannik E. Jakobsen
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  2. Juan Li
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  3. Peter M. Kragh
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  11. Gábor Vajta
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  12. Lars Bolund
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  15. Anders Lade Nielsen
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  16. Jacob Giehm Mikkelsen
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Correspondence to Jacob Giehm Mikkelsen.

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Anders Lade Nielsen and Jacob Giehm Mikkelsen contributed equally to this work.

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Jakobsen, J.E., Li, J., Kragh, P.M. et al. Pig transgenesis by Sleeping Beauty DNA transposition. Transgenic Res 20, 533–545 (2011). https://doi.org/10.1007/s11248-010-9438-x

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