title: Establishment of cell-based transposon-mediated transgenesis in cattle.
abstract: Transposon-mediated transgenesis is a well-established tool for genome modification in small animal models. However, translation of this active transgenic method to large animals warrants further investigations. Here, the piggyBac (PB) and sleeping beauty (SB) transposon systems were assessed for stable gene transfer into the cattle genome. Bovine fibroblasts were transfected either with a helper-independent PB system or a binary SB system. Both transposons were highly active in bovine cells increasing the efficiency of DNA integration up to 88 times over basal nonfacilitated integrations in a colony formation assay. SB transposase catalyzed multiplex transgene integrations in fibroblast cells transfected with the helper vector and two donor vectors carrying different transgenes (fluorophore and neomycin resistance). Stably transfected fibroblasts were used for SCNT and on in vitro embryo culture, morphologically normal blastocysts that expressed the fluorophore were obtained with both transposon systems. The data indicate that transposition is a feasible approach for genetic engineering in the cattle genome.
authors.affiliation: Departamento de Biología Molecular, Facultad de Ciencias Exactas, Fco-Qcas y Naturales, Universidad Nacional de Río Cuarto, Córdoba, República Argentina.
authors.forename: Ana P
authors.initials: AP
authors.lastname: Alessio
authors.affiliation: Departamento de Biología Molecular, Facultad de Ciencias Exactas, Fco-Qcas y Naturales, Universidad Nacional de Río Cuarto, Córdoba, República Argentina.
authors.forename: Alejandro E
authors.initials: AE
authors.lastname: Fili
authors.affiliation: Department of Biotechnology, Friedrich-Loeffler-Institut, Institut für Nutztiergenetik, Neustadt, Germany.
authors.forename: Wiebke
authors.initials: W
authors.lastname: Garrels
authors.affiliation: Departamento de Biología Molecular, Facultad de Ciencias Exactas, Fco-Qcas y Naturales, Universidad Nacional de Río Cuarto, Córdoba, República Argentina.
authors.forename: Diego O
authors.initials: DO
authors.lastname: Forcato
authors.affiliation: Departamento de Biología Molecular, Facultad de Ciencias Exactas, Fco-Qcas y Naturales, Universidad Nacional de Río Cuarto, Córdoba, República Argentina.
authors.forename: María F
authors.initials: MF
authors.lastname: Olmos Nicotra
authors.affiliation: Departamento de Biología Molecular, Facultad de Ciencias Exactas, Fco-Qcas y Naturales, Universidad Nacional de Río Cuarto, Córdoba, República Argentina.
authors.forename: Ana C
authors.initials: AC
authors.lastname: Liaudat
authors.affiliation: Laboratorio de Biotecnología Animal, Departamento de Producción Animal, Facultad de Agronomía, Universidad de Buenos Aires, Buenos Aires, República Argentina.
authors.forename: Romina J
authors.initials: RJ
authors.lastname: Bevacqua
authors.affiliation: Laboratorio de Biotecnología Animal, Departamento de Producción Animal, Facultad de Agronomía, Universidad de Buenos Aires, Buenos Aires, República Argentina.
authors.forename: Virginia
authors.initials: V
authors.lastname: Savy
authors.affiliation: Laboratorio de Biotecnología Animal, Departamento de Producción Animal, Facultad de Agronomía, Universidad de Buenos Aires, Buenos Aires, República Argentina.
authors.forename: María I
authors.initials: MI
authors.lastname: Hiriart
authors.affiliation: Department of Biotechnology, Friedrich-Loeffler-Institut, Institut für Nutztiergenetik, Neustadt, Germany.
authors.forename: Thirumala R
authors.initials: TR
authors.lastname: Talluri
authors.affiliation: Department of Anatomy, Biochemistry and Physiology, Institute for Biogenesis Research, John A. Burns School of Medicine, University of Hawaii at Manoa, Honolulu, Hawaii, USA.
authors.forename: Jesse B
authors.initials: JB
authors.lastname: Owens
authors.affiliation: Division of Medical Biotechnology, Paul-Ehrlich-Institute, Langen, Germany.
authors.forename: Zoltán
authors.initials: Z
authors.lastname: Ivics
authors.affiliation: Laboratorio de Biotecnología Animal, Departamento de Producción Animal, Facultad de Agronomía, Universidad de Buenos Aires, Buenos Aires, República Argentina.
authors.forename: Daniel F
authors.initials: DF
authors.lastname: Salamone
authors.affiliation: Department of Anatomy, Biochemistry and Physiology, Institute for Biogenesis Research, John A. Burns School of Medicine, University of Hawaii at Manoa, Honolulu, Hawaii, USA.
authors.forename: Stefan
authors.initials: S
authors.lastname: Moisyadi
authors.affiliation: Department of Biotechnology, Friedrich-Loeffler-Institut, Institut für Nutztiergenetik, Neustadt, Germany.
authors.forename: Wilfried A
authors.initials: WA
authors.lastname: Kues
authors.affiliation: Departamento de Biología Molecular, Facultad de Ciencias Exactas, Fco-Qcas y Naturales, Universidad Nacional de Río Cuarto, Córdoba, República Argentina. Electronic address: pbosch@exa.unrc.edu.ar.
authors.forename: Pablo
authors.initials: P
authors.lastname: Bosch
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D002417:Cattle / Q000235:genetics*; D002460:Cell Line; D004251:DNA Transposable Elements / Q000235:genetics*; D005347:Fibroblasts; D005822:Genetic Vectors / Q000235:genetics*; D053652:Nuclear Transfer Techniques; D014162:Transfection; D019895:Transposases
chemical_list: D004251:DNA Transposable Elements; D019895:Transposases
keywords: Cattle; PiggyBac; Sleeping beauty; Transgenesis; Transposon