Pig transgenesis by Sleeping Beauty DNA transposition.
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.
Jakobsen
Jannik E
JE
Department of Human Genetics, University of Aarhus, Wilh. Meyers Allé 1240, 8000, Aarhus C, Denmark.
Li
Juan
J
Kragh
Peter M
PM
Moldt
Brian
B
Lin
Lin
L
Liu
Ying
Y
Schmidt
Mette
M
Winther
Kjeld Dahl
KD
Schyth
Brian Dall
BD
Holm
Ida E
IE
Vajta
Gábor
G
Bolund
Lars
L
Callesen
Henrik
H
Jørgensen
Arne Lund
AL
Nielsen
Anders Lade
AL
Mikkelsen
Jacob Giehm
JG
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*
D004251:DNA Transposable Elements; D049452:Green Fluorescent Proteins; D019895:Transposases
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