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Hemizygous minipigs produced by random gene insertion and handmade cloning express the Alzheimer’s disease-causing dominant mutation APPsw

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Abstract

In an effort to develop a porcine model of Alzheimer’s disease we used handmade cloning to produce seven transgenic Göttingen minipigs. The donor fibroblasts had been stably transfected with a plasmid cassette containing, as transgene, the cDNA of the neuronal variant of the human amyloid precursor protein gene with the Swedish mutation preceded by beta-globin sequences to induce splicing and a human PDGFbeta promoter fragment to drive transcription. Transgene insertion had occurred only at the GLIS3 locus where a single complete copy of the transgene was identified in intronic sequences in opposite direction. Similar and robust levels of the transgene transcript were detected in skin biopsies from all piglets and the sequence of full-length transcript was verified. Consistent with PDGFbeta promoter function, high levels of transgene expression, including high level of the corresponding protein, was observed in brain tissue and not in heart or liver tissues. A rough estimate predicts that accumulation of the Aβ peptide in the brain may develop at the age of 1–2 years.

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Abbreviations

APP:

Amyloid precursor protein

APP695sw:

Neuronal splicevariant of APP with the Swedish mutation

Aβ:

Amyloid beta-protein

AD:

Alzheimer’s disease

PSEN:

Presenilin

NFT:

Neurofibrillary tangles

HMC:

Handmade cloning

SCNT:

Somatic cell nuclear transfer

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Acknowledgments

This work was supported financially by the Danish Research Agency Grant no. 274-05-0197, the Danish Agency for Science, Technology, and Innovation Grant no. 271-05-0197, and Institute of Clinical Medicine, University of Aarhus. ALN is supported by a Hallas-Møller Fellowship from the NovoNordisk Foundation.

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Author notes

  1. Ingrid Brück Bøgh

    Present address: , Novo Nordisk A/S, Novo Nordisk Park, 2760, Måløv, Denmark

  2. Gábor Vajta

    Present address: , PIVET Medical Centre, 166-168 Cambridge Street, Leederville, Perth, WA, 6007, Australia

Authors and Affiliations

  1. Department of Human Genetics, Faculty of Health Sciences, The Bartholin Building, Aarhus University, 8000, Aarhus C, Denmark

    Peter M. Kragh, Anders Lade Nielsen, Jannik E. Jakobsen, Marianne G. Johansen, Lars Bolund & Arne Lund Jørgensen

  2. Department of Genetics and Biotechnology, Faculty of Agricultural Sciences, Aarhus University, 8830, Tjele, Denmark

    Peter M. Kragh, Juan Li, Yutao Du, Lin Lin & Gábor Vajta

  3. Department of Veterinary Reproduction and Obstetrics, Faculty of Life Sciences, University of Copenhagen, 1870, Frederiksberg C, Denmark

    Mette Schmidt & Ingrid Brück Bøgh

  4. Department of Pathology, Aalborg Hospital, Aarhus University Hospital, 9000, Aalborg, Denmark

    Ida E. Holm

  5. Department of Animal Health, Welfare and Nutrition, Faculty of Agricultural Sciences, Aarhus University, 8830, Tjele, Denmark

    Stig Purup

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  1. Peter M. Kragh
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Correspondence to Arne Lund Jørgensen.

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P. M. Kragh and A. L. Nielsen shares 1 authorship.

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Kragh, P.M., Nielsen, A.L., Li, J. et al. Hemizygous minipigs produced by random gene insertion and handmade cloning express the Alzheimer’s disease-causing dominant mutation APPsw. Transgenic Res 18, 545–558 (2009). https://doi.org/10.1007/s11248-009-9245-4

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