title: Pig transgenesis by Sleeping Beauty DNA transposition.
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.
authors.affiliation: Department of Human Genetics, University of Aarhus, Wilh. Meyers Allé 1240, 8000, Aarhus C, Denmark.
authors.forename: Jannik E
authors.initials: JE
authors.lastname: Jakobsen
authors.forename: Juan
authors.initials: J
authors.lastname: Li
authors.forename: Peter M
authors.initials: PM
authors.lastname: Kragh
authors.forename: Brian
authors.initials: B
authors.lastname: Moldt
authors.forename: Lin
authors.initials: L
authors.lastname: Lin
authors.forename: Ying
authors.initials: Y
authors.lastname: Liu
authors.forename: Mette
authors.initials: M
authors.lastname: Schmidt
authors.forename: Kjeld Dahl
authors.initials: KD
authors.lastname: Winther
authors.forename: Brian Dall
authors.initials: BD
authors.lastname: Schyth
authors.forename: Ida E
authors.initials: IE
authors.lastname: Holm
authors.forename: Gábor
authors.initials: G
authors.lastname: Vajta
authors.forename: Lars
authors.initials: L
authors.lastname: Bolund
authors.forename: Henrik
authors.initials: H
authors.lastname: Callesen
authors.forename: Arne Lund
authors.initials: AL
authors.lastname: Jørgensen
authors.forename: Anders Lade
authors.initials: AL
authors.lastname: Nielsen
authors.forename: Jacob Giehm
authors.initials: JG
authors.lastname: Mikkelsen
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D004251:DNA Transposable Elements / Q000235:genetics*; D005347:Fibroblasts / Q000378:metabolism; D018014:Gene Transfer Techniques; D005818:Genetic Engineering / Q000379:methods; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D013552:Swine; D013556:Swine, Miniature; D019076:Transgenes / Q000235:genetics / Q000502:physiology; D019895:Transposases / Q000235:genetics / Q000378:metabolism*
chemical_list: D004251:DNA Transposable Elements; D049452:Green Fluorescent Proteins; D019895:Transposases
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