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J Vet Med Sci ; 73(11): 1453-7, 2011 Nov.
Article in English | MEDLINE | ID: mdl-21747215

ABSTRACT

Transgenic research on cattle embryos has been developed to date using viral or plasmid DNA delivery systems. In this study, a different gene delivery system, piggybac transposition, was employed to investigate if it can be applied for producing transgenic cattle embryos. Green or red fluorescent proteins (GFP or RFP) were transfected into donor fibroblasts, and then transfected donor cells were reprogrammed in enucleated oocytes through SCNT and developed into pre-implantation stage embryos. GFP was expressed in donor cells and in cloned embryos without any mosaicism. Induction of RFP expression was regulated by doxycycline treatment in donor fibroblasts and pre-implantational stage embryos. In conclusion, this study demonstrated that piggybac transposition could be a mean to deliver genes into bovine somatic cells or embryos for transgenic research.


Subject(s)
Animals, Genetically Modified , Cattle/genetics , Genetic Engineering/veterinary , Nuclear Transfer Techniques/veterinary , Animals , Cattle/embryology , Doxycycline/metabolism , Fibroblasts , Gene Expression Regulation, Developmental , Genetic Engineering/methods , Green Fluorescent Proteins/genetics , Green Fluorescent Proteins/metabolism , Luminescent Proteins/genetics , Luminescent Proteins/metabolism , Reverse Transcriptase Polymerase Chain Reaction/veterinary , Transfection/methods , Transfection/veterinary , Red Fluorescent Protein
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