Hemizygous minipigs produced by random gene insertion and handmade cloning express the Alzheimer's disease-causing dominant mutation APPsw.
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 PDGF beta 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 PDGF beta 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 A beta peptide in the brain may develop at the age of 1-2 years.
Kragh
Peter M
PM
Department of Human Genetics, Faculty of Health Sciences, The Bartholin Building, Aarhus University, 8000, Aarhus C, Denmark.
Nielsen
Anders Lade
AL
Li
Juan
J
Du
Yutao
Y
Lin
Lin
L
Schmidt
Mette
M
Bøgh
Ingrid Brück
IB
Holm
Ida E
IE
Jakobsen
Jannik E
JE
Johansen
Marianne G
MG
Purup
Stig
S
Bolund
Lars
L
Vajta
Gábor
G
Jørgensen
Arne Lund
AL
D000544:Alzheimer Disease / Q000235:genetics*; D016229:Amyloid beta-Peptides / Q000378:metabolism; D016564:Amyloid beta-Protein Precursor / Q000235:genetics*; D000818:Animals; D030801:Animals, Genetically Modified; D001921:Brain / Q000378:metabolism; D003001:Cloning, Molecular; D005347:Fibroblasts / Q000378:metabolism; D005799:Genes, Dominant; D006801:Humans; D016254:Mutagenesis, Insertional; D009154:Mutation; D012326:RNA Splicing; D020797:Receptor, Platelet-Derived Growth Factor beta / Q000235:genetics; D013552:Swine; D013556:Swine, Miniature / Q000235:genetics*; D014162:Transfection; D019076:Transgenes
D016229:Amyloid beta-Peptides; D016564:Amyloid beta-Protein Precursor; D020797:Receptor, Platelet-Derived Growth Factor beta