Pig transgenesis by Sleeping Beauty DNA transposition

Publication: Research - peer-reviewJournal article – Annual report year: 2010

  • Author: Jakobsen, Jannik E.

    Aarhus University

  • Author: Li, Juan

    Aarhus University

  • Author: Kragh, Peter M.

    Aarhus University

  • Author: Moldt, Brian

    Aarhus University

  • Author: Lin, Lin

    Aarhus University

  • Author: Liu, Ying

    Aarhus University

  • Author: Schmidt, Mette

    University of Copenhagen

  • Author: Winther, Kjeld D.

    Danish Agriculture and Food Council

  • Author: Schyth, Brian Dall

    Section of Fish Diseases, Division of Poultry, Fish and Fur Animals, National Veterinary Institute, Technical University of Denmark

  • Author: Holm, Ida E.

    Randers Hospital, Department of Pathology

  • Author: Vajta, Gabor

    Aarhus University

  • Author: Bolund, Lars

    Aarhus University

  • Author: Callesen, Henrik

    Aarhus University

  • Author: Jørgensen, Arne L.

    Aarhus University

  • Author: Nielsen, Anders L.

    Aarhus University

  • Author: Mikkelsen, Jacob G.

    Aarhus University

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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.
Original languageEnglish
JournalTransgenic Research
Publication date2011
Volume20
Issue3
Pages533-545
ISSN0962-8819
DOIs
StatePublished

Bibliographical note

The original publication is available at www.springerlink.com

CitationsWeb of Science® Times Cited: 25

Keywords

  • Systemic gene expression, Pig transgenesis, Genetic engineering, Somatic cell nuclear transfer, Sleeping beauty DNA transposition, DNA transposase
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