Journal article
Pig transgenesis by Sleeping Beauty DNA transposition
Aarhus University1
University of Copenhagen2
Danish Agriculture and Food Council3
Section of Fish Diseases, Division of Poultry, Fish and Fur Animals, National Veterinary Institute, Technical University of Denmark4
Division of Poultry, Fish and Fur Animals, National Veterinary Institute, Technical University of Denmark5
National Veterinary Institute, Technical University of Denmark6
Region Hospital Randers7
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.
Language: | English |
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Publisher: | Springer Netherlands |
Year: | 2011 |
Pages: | 533-545 |
Journal subtitle: | Associated With the International Society for Transgenic Technologies (istt) |
ISSN: | 15739368 and 09628819 |
Types: | Journal article |
DOI: | 10.1007/s11248-010-9438-x |
ORCIDs: | 0000-0002-7546-4948 , 0000-0002-0185-7300 , 0000-0003-4372-9961 and 0000-0002-1322-3209 |
DNA transposase Genetic engineering Pig transgenesis SDG 3 - Good Health and Well-being Sleeping beauty DNA transposition Somatic cell nuclear transfer Systemic gene expression
Animal Genetics and Genomics Animals Animals, Genetically Modified Base Sequence Biomedicine DNA Transposable Elements Fibroblasts Gene Transfer Techniques Genetic Engineering Green Fluorescent Proteins Human Genetics Humans Molecular Medicine Molecular Sequence Data Mutagenesis, Insertional Plant Genetics & Genomics Plant Sciences Swine Swine, Miniature Transgenes Transposases