Establishment of cell-based transposon-mediated transgenesis in cattle
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 st...
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todo:paper_0093691X_v85_n7_p1297_Alessio2023-10-03T14:55:53Z Establishment of cell-based transposon-mediated transgenesis in cattle Alessio, A.P. Fili, A.E. Garrels, W. Forcato, D.O. Olmos Nicotra, M.F. Liaudat, A.C. Bevacqua, R.J. Savy, V. Hiriart, M.I. Talluri, T.R. Owens, J.B. Ivics, Z. Salamone, D.F. Moisyadi, S. Kues, W.A. Bosch, P. Cattle PiggyBac Sleeping beauty Transgenesis Transposon transposase transposon animal bovine cell line cell nucleus transplantation fibroblast gene vector genetic transfection genetics transgenic animal transposon Animals Animals, Genetically Modified Cattle Cell Line DNA Transposable Elements Fibroblasts Genetic Vectors Nuclear Transfer Techniques Transfection Transposases 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. © 2016 Elsevier Inc. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_0093691X_v85_n7_p1297_Alessio |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
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Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Cattle PiggyBac Sleeping beauty Transgenesis Transposon transposase transposon animal bovine cell line cell nucleus transplantation fibroblast gene vector genetic transfection genetics transgenic animal transposon Animals Animals, Genetically Modified Cattle Cell Line DNA Transposable Elements Fibroblasts Genetic Vectors Nuclear Transfer Techniques Transfection Transposases |
| spellingShingle |
Cattle PiggyBac Sleeping beauty Transgenesis Transposon transposase transposon animal bovine cell line cell nucleus transplantation fibroblast gene vector genetic transfection genetics transgenic animal transposon Animals Animals, Genetically Modified Cattle Cell Line DNA Transposable Elements Fibroblasts Genetic Vectors Nuclear Transfer Techniques Transfection Transposases Alessio, A.P. Fili, A.E. Garrels, W. Forcato, D.O. Olmos Nicotra, M.F. Liaudat, A.C. Bevacqua, R.J. Savy, V. Hiriart, M.I. Talluri, T.R. Owens, J.B. Ivics, Z. Salamone, D.F. Moisyadi, S. Kues, W.A. Bosch, P. Establishment of cell-based transposon-mediated transgenesis in cattle |
| topic_facet |
Cattle PiggyBac Sleeping beauty Transgenesis Transposon transposase transposon animal bovine cell line cell nucleus transplantation fibroblast gene vector genetic transfection genetics transgenic animal transposon Animals Animals, Genetically Modified Cattle Cell Line DNA Transposable Elements Fibroblasts Genetic Vectors Nuclear Transfer Techniques Transfection Transposases |
| description |
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. © 2016 Elsevier Inc. |
| format |
JOUR |
| author |
Alessio, A.P. Fili, A.E. Garrels, W. Forcato, D.O. Olmos Nicotra, M.F. Liaudat, A.C. Bevacqua, R.J. Savy, V. Hiriart, M.I. Talluri, T.R. Owens, J.B. Ivics, Z. Salamone, D.F. Moisyadi, S. Kues, W.A. Bosch, P. |
| author_facet |
Alessio, A.P. Fili, A.E. Garrels, W. Forcato, D.O. Olmos Nicotra, M.F. Liaudat, A.C. Bevacqua, R.J. Savy, V. Hiriart, M.I. Talluri, T.R. Owens, J.B. Ivics, Z. Salamone, D.F. Moisyadi, S. Kues, W.A. Bosch, P. |
| author_sort |
Alessio, A.P. |
| title |
Establishment of cell-based transposon-mediated transgenesis in cattle |
| title_short |
Establishment of cell-based transposon-mediated transgenesis in cattle |
| title_full |
Establishment of cell-based transposon-mediated transgenesis in cattle |
| title_fullStr |
Establishment of cell-based transposon-mediated transgenesis in cattle |
| title_full_unstemmed |
Establishment of cell-based transposon-mediated transgenesis in cattle |
| title_sort |
establishment of cell-based transposon-mediated transgenesis in cattle |
| url |
http://hdl.handle.net/20.500.12110/paper_0093691X_v85_n7_p1297_Alessio |
| work_keys_str_mv |
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1807323645197942784 |