Pig transgenesis by Sleeping Beauty DNA transposition

Jannik E. Jakobsen, Juan Li, Peter M. Kragh, Brian Moldt, Lin Lin, Ying Liu, Mette Schmidt, Kjeld D. Winther, Brian Dall Schyth, Ida E. Holm, Gabor Vajta, Lars Bolund, Henrik Callesen, Arne L. Jørgensen, Anders L. Nielsen, Jacob G. Mikkelsen

    Research output: Contribution to journalJournal articleResearchpeer-review

    520 Downloads (Pure)

    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.
    Original languageEnglish
    JournalTransgenic Research
    Volume20
    Issue number3
    Pages (from-to)533-545
    ISSN0962-8819
    DOIs
    Publication statusPublished - 2011

    Bibliographical note

    The original publication is available at www.springerlink.com

    Keywords

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

    Fingerprint Dive into the research topics of 'Pig transgenesis by Sleeping Beauty DNA transposition'. Together they form a unique fingerprint.

    Cite this