Pig transgenesis by Sleeping Beauty DNA transposition

Jannik E. Jakobsen; Juan Li; Peter M Kragh; Brian Moldt; Lin Lin; Ying Liu; Mette Schmidt; Kjeld Dahl Winther; Brian Dall Schyth; Ida Elisabeth Holm; Gabor Vajta; Lars Bolund; Henrik Callesen; Arne Lund Jørgensen; Anders Lade Nielsen; Jacob Giehm Mikkelsen

doi:10.1007/s11248-010-9438-x

Pig transgenesis by Sleeping Beauty DNA transposition

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

Department of Animal and Veterinary Sciences - Reproductive Biology

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

54 Citations (Scopus)

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 language	English
Journal	Transgenic Research
Volume	20
Issue	3
Pages (from-to)	533-545
Number of pages	13
ISSN	0962-8819
DOIs	https://doi.org/10.1007/s11248-010-9438-x
Publication status	Published - 2011

Keywords

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

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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. G{\"o}ttingen 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.",

keywords = "Pig transgenesis, Somatic cell nuclear transfer, Genetic engineering, Sleeping beauty DNA transposition, DNA transposase, Systemic gene expression",

author = "Jakobsen, {Jannik E.} and Juan Li and Kragh, {Peter M} and Brian Moldt and Lin Lin and Ying Liu and Mette Schmidt and Winther, {Kjeld Dahl} and Schyth, {Brian Dall} and Holm, {Ida Elisabeth} and Gabor Vajta and Lars Bolund and Henrik Callesen and J{\o}rgensen, {Arne Lund} and Nielsen, {Anders Lade} and Mikkelsen, {Jacob Giehm}",

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AU - Moldt, Brian

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AU - Liu, Ying

AU - Schmidt, Mette

AU - Winther, Kjeld Dahl

AU - Schyth, Brian Dall

AU - Holm, Ida Elisabeth

AU - Vajta, Gabor

AU - Bolund, Lars

AU - Callesen, Henrik

AU - Jørgensen, Arne Lund

AU - Nielsen, Anders Lade

AU - Mikkelsen, Jacob Giehm

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AB - 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.

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KW - Somatic cell nuclear transfer

KW - Genetic engineering

KW - Sleeping beauty DNA transposition

KW - DNA transposase

KW - Systemic gene expression

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JO - Transgenic Research

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Pig transgenesis by Sleeping Beauty DNA transposition

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Keywords

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