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Germline transgenesis in pigs by cytoplasmic microinjection of Sleeping Beauty transposons
Z. Ivics, W. Garrels, L. Mátés, TY. Yau, S. Bashir, V. Zidek, V. Landa, A. Geurts, M. Pravenec, T. Rülicke, WA. Kues, Z. Izsvák,
Language English Country England, Great Britain
Document type Journal Article, Research Support, Non-U.S. Gov't
NLK
ProQuest Central
from 2006-06-01 to 1 year ago
Medline Complete (EBSCOhost)
from 2013-03-01 to 2015-12-31
Health & Medicine (ProQuest)
from 2006-06-01 to 1 year ago
- MeSH
- Genetic Vectors MeSH
- Animals, Genetically Modified * MeSH
- Genome MeSH
- Microinjections MeSH
- Swine genetics MeSH
- Gene Transfer Techniques * MeSH
- Transposases MeSH
- DNA Transposable Elements * MeSH
- Germ Cells MeSH
- Animals MeSH
- Check Tag
- Male MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
The pig has emerged as an important large animal model in biomedical and pharmaceutical research. We describe a protocol for high-efficiency germline transgenesis and sustained transgene expression in pigs by using the Sleeping Beauty (SB) transposon system. The protocol is based on co-injection of a plasmid encoding the SB100X hyperactive transposase, together with a second plasmid carrying a transgene flanked by binding sites for the transposase, into the cytoplasm of porcine zygotes. The transposase mediates excision of the transgene cassette from the plasmid vector and its permanent insertion into the genome to produce stable transgenic animals. This method compares favorably in terms of both efficiency and reliable transgene expression to classic pronuclear microinjection or somatic cell nuclear transfer (SCNT), and it offers comparable efficacies to lentiviral approaches, without limitations on vector design, issues of transgene silencing and the toxicity and biosafety concerns of working with viral vectors. Microinjection of the vectors into zygotes and transfer of the embryos to recipient animals can be performed in 1 d; generation of germline-transgenic lines by using this protocol takes ∼1 year.
] Division of Medical Biotechnology Paul Ehrlich Institute Langen Germany [2]
] Friedrich Loeffler Institut Institut für Nutztiergenetik Neustadt Germany [2]
Biological Research Centre Hungarian Academy of Sciences Szeged Hungary
Department of Physiology Medical College of Wisconsin Milwaukee Wisconsin USA
Friedrich Loeffler Institut Institut für Nutztiergenetik Neustadt Germany
Institute of Laboratory Animal Science University of Veterinary Medicine Vienna Vienna Austria
Institute of Physiology Academy of Sciences of the Czech Republic Prague Czech Republic
References provided by Crossref.org
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