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dc.contributor.author
Vichera, Gabriel Damian
dc.contributor.author
Moro, Lucía Natalia
dc.contributor.author
Salamone, Daniel Felipe
dc.date.available
2017-05-02T20:56:25Z
dc.date.issued
2011-04
dc.identifier.citation
Vichera, Gabriel Damian; Moro, Lucía Natalia; Salamone, Daniel Felipe; Efficient transgene expression in IVF and parthenogenetic bovine embryos by intracytoplasmic injection of DNA-liposome complexes; Wiley; Reproduction In Domestic Animals; 46; 2; 4-2011; 214-220
dc.identifier.issn
0936-6768
dc.identifier.uri
http://hdl.handle.net/11336/15902
dc.description.abstract
Transgenic animals constitute an important tool with many biotechnological applications. Although there have been advances in this field, we propose a novel method that may greatly increase the efficiency of transgenic animal production and thereby its application. This new technique consists of intracytoplasmic injection of liposomes, in bovine oocytes and zygotes, to introduce exogenous DNA. In the first experiment, we evaluated embryo development and EGFP expression in In Vitro Fertilization (IVF) embryos injected with different concentrations of exogenous DNA-liposome complexes (0.5, 5, 50, 500 ng pCX-EGFP/μl). The highest EGFP-embryos rates were obtained using 500 ng pCX-EGFP/μl. In the second experiment, we evaluated embryo development and EGFP expression following the injection of DNA-liposome complexes into pre-fertilized oocytes and presumptive zygotes, 16 and 24 h post-fertilization. Approximately 70% of the cleaved embryos and 50% of the blastocysts expressed EGFP, when egfp-liposome was injected 16 h post-fertilization. The percentages of positive embryos for the 24-h post-fertilization and pre-fertilization groups were 30.1 and 6.3, respectively. Blastocysts that developed from injected zygotes were analysed by PCR, confirming the presence of transgene in all embryos. Finally, we examined the embryo development and EGFP expression of parthenogenetic embryos that resulted from the injection of egfp-liposome complexes into pre-activated oocytes, and 3 and 11 h post-activated oocytes. The group with the highest expression rate (48.4%) was the one injected 3 h post-activation. In summary, this study reports the efficient, reproducible and fast production of IVF and parthenogenetic embryos expressing EGFP, by the intracytoplasmic injection of liposomes to introduce the foreign DNA.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Wiley
dc.rights
info:eu-repo/semantics/restrictedAccess
dc.rights.uri
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.subject
Ntracytoplasmic Injection of Dna-Liposome Complexes
dc.subject
Transgene Expression
dc.subject.classification
Tecnología GM, clonación de ganado, selección asistida, diagnósticos, tecnología de producción de biomasa, etc.
dc.subject.classification
Biotecnología Agropecuaria
dc.subject.classification
CIENCIAS AGRÍCOLAS
dc.title
Efficient transgene expression in IVF and parthenogenetic bovine embryos by intracytoplasmic injection of DNA-liposome complexes
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2017-04-24T20:43:31Z
dc.journal.volume
46
dc.journal.number
2
dc.journal.pagination
214-220
dc.journal.pais
Reino Unido
dc.journal.ciudad
Londres
dc.description.fil
Fil: Vichera, Gabriel Damian. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
dc.description.fil
Fil: Moro, Lucía Natalia. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
dc.description.fil
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
dc.journal.title
Reproduction In Domestic Animals
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.1111/j.1439-0531.2010.01642.x
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/http://onlinelibrary.wiley.com/doi/10.1111/j.1439-0531.2010.01642.x/abstract


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