Assessing Tn5 and sleeping beauty for transpositional transgenesis by cytoplasmic injection into bovine and ovine zygotes
Transgenic domestic animals represent an alternative to bioreactors for large-scale production of biopharmaceuticals and could also provide more accurate biomedical models than rodents. However, their generation remains inefficient. Recently, DNA transposons allowed improved transgenesis efficiencie...
| Autores principales: | , , , , , , , , , , , , , , , |
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| Formato: | Artículo |
| Lenguaje: | Inglés |
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2017
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| Acceso en línea: | http://hdl.handle.net/20.500.12123/559 http://journals.plos.org/plosone/article/related?id=10.1371/journal.pone.0174025 https://doi.org/10.1371/journal.pone.0174025 |
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| author | Bevacqua, Romina Jimena Fernández y Martin, Rafael Canel, Natalia Gabriela Gibbons, Alejandro Eduardo Texeira, D.I.A. Lange, F. Vans Landschoot, Geraldina Savy, Virginia Briski, Olinda Hiriart, María Inés Grueso, Esther Ivics, Zoltán Taboga, Oscar Alberto Kues, Wilfried A. Ferraris, S.R. Salamone, Daniel Felipe |
| author_browse | Bevacqua, Romina Jimena Briski, Olinda Canel, Natalia Gabriela Fernández y Martin, Rafael Ferraris, S.R. Gibbons, Alejandro Eduardo Grueso, Esther Hiriart, María Inés Ivics, Zoltán Kues, Wilfried A. Lange, F. Salamone, Daniel Felipe Savy, Virginia Taboga, Oscar Alberto Texeira, D.I.A. Vans Landschoot, Geraldina |
| author_facet | Bevacqua, Romina Jimena Fernández y Martin, Rafael Canel, Natalia Gabriela Gibbons, Alejandro Eduardo Texeira, D.I.A. Lange, F. Vans Landschoot, Geraldina Savy, Virginia Briski, Olinda Hiriart, María Inés Grueso, Esther Ivics, Zoltán Taboga, Oscar Alberto Kues, Wilfried A. Ferraris, S.R. Salamone, Daniel Felipe |
| author_sort | Bevacqua, Romina Jimena |
| collection | INTA Digital |
| description | Transgenic domestic animals represent an alternative to bioreactors for large-scale production of biopharmaceuticals and could also provide more accurate biomedical models than rodents. However, their generation remains inefficient. Recently, DNA transposons allowed improved transgenesis efficiencies in mice and pigs. In this work, Tn5 and Sleeping Beauty (SB) transposon systems were evaluated for transgenesis by simple cytoplasmic injection in livestock zygotes. In the case of Tn5, the transposome complex of transposon nucleic acid and Tn5 protein was injected. In the case of SB, the supercoiled plasmids encoding a transposon and the SB transposase were co-injected. In vitro produced bovine zygotes were used to establish the cytoplasmic injection conditions. The in vitro cultured blastocysts were evaluated for reporter gene expression and genotyped. Subsequently, both transposon systems were injected in seasonally available ovine zygotes, employing transposons carrying the recombinant human factor IX driven by the beta-lactoglobulin promoter. The Tn5 approach did not result in transgenic lambs. In contrast, the Sleeping Beauty injection resulted
in 2 lambs (29%) carrying the transgene. Both animals exhibited cellular mosaicism of the transgene. The extraembryonic tissues (placenta or umbilical cord) of three additional animals were also transgenic. These results show that transpositional transgenesis by cytoplasmic injection of SB transposon components can be applied for the production of transgenic lambs of pharmaceutical interest. |
| format | Artículo |
| id | INTA559 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2017 |
| publishDateRange | 2017 |
| publishDateSort | 2017 |
| record_format | dspace |
| spelling | INTA5592018-01-12T13:37:11Z Assessing Tn5 and sleeping beauty for transpositional transgenesis by cytoplasmic injection into bovine and ovine zygotes Bevacqua, Romina Jimena Fernández y Martin, Rafael Canel, Natalia Gabriela Gibbons, Alejandro Eduardo Texeira, D.I.A. Lange, F. Vans Landschoot, Geraldina Savy, Virginia Briski, Olinda Hiriart, María Inés Grueso, Esther Ivics, Zoltán Taboga, Oscar Alberto Kues, Wilfried A. Ferraris, S.R. Salamone, Daniel Felipe Ovinos Bovinae Cigotos Transferencia de Genes Inyección Brucelosis Sheep Zygotes Gene Transfer Injection Tn5 Transgenic domestic animals represent an alternative to bioreactors for large-scale production of biopharmaceuticals and could also provide more accurate biomedical models than rodents. However, their generation remains inefficient. Recently, DNA transposons allowed improved transgenesis efficiencies in mice and pigs. In this work, Tn5 and Sleeping Beauty (SB) transposon systems were evaluated for transgenesis by simple cytoplasmic injection in livestock zygotes. In the case of Tn5, the transposome complex of transposon nucleic acid and Tn5 protein was injected. In the case of SB, the supercoiled plasmids encoding a transposon and the SB transposase were co-injected. In vitro produced bovine zygotes were used to establish the cytoplasmic injection conditions. The in vitro cultured blastocysts were evaluated for reporter gene expression and genotyped. Subsequently, both transposon systems were injected in seasonally available ovine zygotes, employing transposons carrying the recombinant human factor IX driven by the beta-lactoglobulin promoter. The Tn5 approach did not result in transgenic lambs. In contrast, the Sleeping Beauty injection resulted in 2 lambs (29%) carrying the transgene. Both animals exhibited cellular mosaicism of the transgene. The extraembryonic tissues (placenta or umbilical cord) of three additional animals were also transgenic. These results show that transpositional transgenesis by cytoplasmic injection of SB transposon components can be applied for the production of transgenic lambs of pharmaceutical interest. Instituto de Biotecnología Fil: Bevacqua, Romina Jimena. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Fernández y Martín, Rafael. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Canel, Natalia Gabriela. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario; Argentina Fil: Gibbons, Alejandro Eduardo. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Bariloche; Argentina Fil: Texeira, D.I.A. Universidade Estadual do Ceará. Faculdade de Veterinária; Brasil Fil: Lange, F. Universidad Maimónides. Laboratorio de Clonación y Transgenesis; Argentina Fil: Vans Landschoot, Geraldina. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Fil: Lange, F. Universidad Maimónides. Laboratorio de Clonación y Transgenesis; Argentina Fil: Savy, Virginia. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Briski, Olinda. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Hiriart, María Inés. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Grueso, Esther. Paul-Ehrlich-Institute; Alemania Fil: Ivics, Zoltán. Paul-Ehrlich-Institute; Alemania Fil: Taboga, Oscar Alberto. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Fil: Kues, Wilfried A. Friedrich-Loeffler-Institut; Alemania Fil: Ferraris, S.R. Universidad Maimónides. Laboratorio de Clonación y Transgenesis; Argentina Fil: Salamone, Daniel Felipe. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina 2017-07-05T13:25:14Z 2017-07-05T13:25:14Z 2017-03-16 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/559 http://journals.plos.org/plosone/article/related?id=10.1371/journal.pone.0174025 https://doi.org/10.1371/journal.pone.0174025 eng info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) application/pdf PLoS ONE 12 (3) : e0174025 |
| spellingShingle | Ovinos Bovinae Cigotos Transferencia de Genes Inyección Brucelosis Sheep Zygotes Gene Transfer Injection Tn5 Bevacqua, Romina Jimena Fernández y Martin, Rafael Canel, Natalia Gabriela Gibbons, Alejandro Eduardo Texeira, D.I.A. Lange, F. Vans Landschoot, Geraldina Savy, Virginia Briski, Olinda Hiriart, María Inés Grueso, Esther Ivics, Zoltán Taboga, Oscar Alberto Kues, Wilfried A. Ferraris, S.R. Salamone, Daniel Felipe Assessing Tn5 and sleeping beauty for transpositional transgenesis by cytoplasmic injection into bovine and ovine zygotes |
| title | Assessing Tn5 and sleeping beauty for transpositional transgenesis by cytoplasmic injection into bovine and ovine zygotes |
| title_full | Assessing Tn5 and sleeping beauty for transpositional transgenesis by cytoplasmic injection into bovine and ovine zygotes |
| title_fullStr | Assessing Tn5 and sleeping beauty for transpositional transgenesis by cytoplasmic injection into bovine and ovine zygotes |
| title_full_unstemmed | Assessing Tn5 and sleeping beauty for transpositional transgenesis by cytoplasmic injection into bovine and ovine zygotes |
| title_short | Assessing Tn5 and sleeping beauty for transpositional transgenesis by cytoplasmic injection into bovine and ovine zygotes |
| title_sort | assessing tn5 and sleeping beauty for transpositional transgenesis by cytoplasmic injection into bovine and ovine zygotes |
| topic | Ovinos Bovinae Cigotos Transferencia de Genes Inyección Brucelosis Sheep Zygotes Gene Transfer Injection Tn5 |
| url | http://hdl.handle.net/20.500.12123/559 http://journals.plos.org/plosone/article/related?id=10.1371/journal.pone.0174025 https://doi.org/10.1371/journal.pone.0174025 |
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