A unique method to produce transgenic embryos in ovine, porcine, feline, bovine and equine species

Autores
Pereyra Bonnet, Federico Alberto; Fernández y Martín, Rafael; Olivera, R.; Jarazo, J.; Vichera, Gabriel Damian; Gibbons, Alejandro Eduardo; Salamone, Daniel Felipe
Año de publicación
2008
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Transgenesis is an essential tool in many biotechnological applications. Intracytoplasmic sperm injection (ICSI)-mediated gene transfer is a powerful technique to obtain transgenic pups; however, most domestic animal embryos do not develop properly after ICSI. An additional step in the protocol, namely assistance by haploid chemical activation, permits the use of ICSI-mediated gene transfer to generate transgenic preimplantation embryos in a wide range of domestic species, including ovine, porcine, feline, equine and bovine. In the present study, spermatozoa from five species were coincubated with pCX-EGFP plasmid and injected into metaphase II oocytes. The chemical activation protocol consisted of ionomycin plus 6-dimethylaminopurine. We detected high proportions of fluorescent EGFP embryos for all five species (23-60%), but with a high frequency of mosaic expression (range 60-85%). To our knowledge, this is the first study to produce exogenous DNA expression in feline and equine embryos. Chemical activation reduces the lag phase of egfp expression in ovine embryos. Our results show that this unique method could be used to obtain ovine, porcine, feline, bovine and equine transgenic preimplantation embryos. © CSIRO 2008.
Fil: Pereyra Bonnet, Federico Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; Argentina
Fil: Fernández y Martín, Rafael. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; Argentina
Fil: Olivera, R.. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; Argentina
Fil: Jarazo, J.. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; Argentina
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
Fil: Gibbons, Alejandro Eduardo. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Patagonia Norte. Estación Experimental Agropecuaria San Carlos de Bariloche; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte; 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
Materia
CHEMICAL ACTIVATION
GENE EXPRESSION
SPERM MEDIATED
TRANSGENIC MAMMALS
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/130554
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/130554
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling A unique method to produce transgenic embryos in ovine, porcine, feline, bovine and equine speciesPereyra Bonnet, Federico AlbertoFernández y Martín, RafaelOlivera, R.Jarazo, J.Vichera, Gabriel DamianGibbons, Alejandro EduardoSalamone, Daniel FelipeCHEMICAL ACTIVATIONGENE EXPRESSIONSPERM MEDIATEDTRANSGENIC MAMMALShttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Transgenesis is an essential tool in many biotechnological applications. Intracytoplasmic sperm injection (ICSI)-mediated gene transfer is a powerful technique to obtain transgenic pups; however, most domestic animal embryos do not develop properly after ICSI. An additional step in the protocol, namely assistance by haploid chemical activation, permits the use of ICSI-mediated gene transfer to generate transgenic preimplantation embryos in a wide range of domestic species, including ovine, porcine, feline, equine and bovine. In the present study, spermatozoa from five species were coincubated with pCX-EGFP plasmid and injected into metaphase II oocytes. The chemical activation protocol consisted of ionomycin plus 6-dimethylaminopurine. We detected high proportions of fluorescent EGFP embryos for all five species (23-60%), but with a high frequency of mosaic expression (range 60-85%). To our knowledge, this is the first study to produce exogenous DNA expression in feline and equine embryos. Chemical activation reduces the lag phase of egfp expression in ovine embryos. Our results show that this unique method could be used to obtain ovine, porcine, feline, bovine and equine transgenic preimplantation embryos. © CSIRO 2008.Fil: Pereyra Bonnet, Federico Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; ArgentinaFil: Fernández y Martín, Rafael. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; ArgentinaFil: Olivera, R.. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; ArgentinaFil: Jarazo, J.. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; ArgentinaFil: 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; ArgentinaFil: Gibbons, Alejandro Eduardo. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Patagonia Norte. Estación Experimental Agropecuaria San Carlos de Bariloche; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte; ArgentinaFil: 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; ArgentinaCsiro Publishing2008-12info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/130554Pereyra Bonnet, Federico Alberto; Fernández y Martín, Rafael; Olivera, R.; Jarazo, J.; Vichera, Gabriel Damian; et al.; A unique method to produce transgenic embryos in ovine, porcine, feline, bovine and equine species; Csiro Publishing; Reproduction Fertility and Development; 20; 7; 12-2008; 741-7491031-3613CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.publish.csiro.au/rd/rd07172info:eu-repo/semantics/altIdentifier/doi/10.1071/RD07172info:eu-repo/semantics/altIdentifier/url/http://www.provino.com.ar/reproduccion-r/366-a-unique-method-to-produce-transgenic-embryos-in-ovine-porcine-feline-bovine-and-equine-speciesinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:27:38Zoai:ri.conicet.gov.ar:11336/130554instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 10:27:38.743CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv A unique method to produce transgenic embryos in ovine, porcine, feline, bovine and equine species
title A unique method to produce transgenic embryos in ovine, porcine, feline, bovine and equine species
spellingShingle A unique method to produce transgenic embryos in ovine, porcine, feline, bovine and equine species
Pereyra Bonnet, Federico Alberto
CHEMICAL ACTIVATION
GENE EXPRESSION
SPERM MEDIATED
TRANSGENIC MAMMALS
title_short A unique method to produce transgenic embryos in ovine, porcine, feline, bovine and equine species
title_full A unique method to produce transgenic embryos in ovine, porcine, feline, bovine and equine species
title_fullStr A unique method to produce transgenic embryos in ovine, porcine, feline, bovine and equine species
title_full_unstemmed A unique method to produce transgenic embryos in ovine, porcine, feline, bovine and equine species
title_sort A unique method to produce transgenic embryos in ovine, porcine, feline, bovine and equine species
dc.creator.none.fl_str_mv Pereyra Bonnet, Federico Alberto
Fernández y Martín, Rafael
Olivera, R.
Jarazo, J.
Vichera, Gabriel Damian
Gibbons, Alejandro Eduardo
Salamone, Daniel Felipe
author Pereyra Bonnet, Federico Alberto
author_facet Pereyra Bonnet, Federico Alberto
Fernández y Martín, Rafael
Olivera, R.
Jarazo, J.
Vichera, Gabriel Damian
Gibbons, Alejandro Eduardo
Salamone, Daniel Felipe
author_role author
author2 Fernández y Martín, Rafael
Olivera, R.
Jarazo, J.
Vichera, Gabriel Damian
Gibbons, Alejandro Eduardo
Salamone, Daniel Felipe
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv CHEMICAL ACTIVATION
GENE EXPRESSION
SPERM MEDIATED
TRANSGENIC MAMMALS
topic CHEMICAL ACTIVATION
GENE EXPRESSION
SPERM MEDIATED
TRANSGENIC MAMMALS
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Transgenesis is an essential tool in many biotechnological applications. Intracytoplasmic sperm injection (ICSI)-mediated gene transfer is a powerful technique to obtain transgenic pups; however, most domestic animal embryos do not develop properly after ICSI. An additional step in the protocol, namely assistance by haploid chemical activation, permits the use of ICSI-mediated gene transfer to generate transgenic preimplantation embryos in a wide range of domestic species, including ovine, porcine, feline, equine and bovine. In the present study, spermatozoa from five species were coincubated with pCX-EGFP plasmid and injected into metaphase II oocytes. The chemical activation protocol consisted of ionomycin plus 6-dimethylaminopurine. We detected high proportions of fluorescent EGFP embryos for all five species (23-60%), but with a high frequency of mosaic expression (range 60-85%). To our knowledge, this is the first study to produce exogenous DNA expression in feline and equine embryos. Chemical activation reduces the lag phase of egfp expression in ovine embryos. Our results show that this unique method could be used to obtain ovine, porcine, feline, bovine and equine transgenic preimplantation embryos. © CSIRO 2008.
Fil: Pereyra Bonnet, Federico Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; Argentina
Fil: Fernández y Martín, Rafael. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; Argentina
Fil: Olivera, R.. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; Argentina
Fil: Jarazo, J.. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; Argentina
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
Fil: Gibbons, Alejandro Eduardo. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Patagonia Norte. Estación Experimental Agropecuaria San Carlos de Bariloche; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte; 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
description Transgenesis is an essential tool in many biotechnological applications. Intracytoplasmic sperm injection (ICSI)-mediated gene transfer is a powerful technique to obtain transgenic pups; however, most domestic animal embryos do not develop properly after ICSI. An additional step in the protocol, namely assistance by haploid chemical activation, permits the use of ICSI-mediated gene transfer to generate transgenic preimplantation embryos in a wide range of domestic species, including ovine, porcine, feline, equine and bovine. In the present study, spermatozoa from five species were coincubated with pCX-EGFP plasmid and injected into metaphase II oocytes. The chemical activation protocol consisted of ionomycin plus 6-dimethylaminopurine. We detected high proportions of fluorescent EGFP embryos for all five species (23-60%), but with a high frequency of mosaic expression (range 60-85%). To our knowledge, this is the first study to produce exogenous DNA expression in feline and equine embryos. Chemical activation reduces the lag phase of egfp expression in ovine embryos. Our results show that this unique method could be used to obtain ovine, porcine, feline, bovine and equine transgenic preimplantation embryos. © CSIRO 2008.
publishDate 2008
dc.date.none.fl_str_mv 2008-12
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/130554
Pereyra Bonnet, Federico Alberto; Fernández y Martín, Rafael; Olivera, R.; Jarazo, J.; Vichera, Gabriel Damian; et al.; A unique method to produce transgenic embryos in ovine, porcine, feline, bovine and equine species; Csiro Publishing; Reproduction Fertility and Development; 20; 7; 12-2008; 741-749
1031-3613
CONICET Digital
CONICET
url http://hdl.handle.net/11336/130554
identifier_str_mv Pereyra Bonnet, Federico Alberto; Fernández y Martín, Rafael; Olivera, R.; Jarazo, J.; Vichera, Gabriel Damian; et al.; A unique method to produce transgenic embryos in ovine, porcine, feline, bovine and equine species; Csiro Publishing; Reproduction Fertility and Development; 20; 7; 12-2008; 741-749
1031-3613
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.publish.csiro.au/rd/rd07172
info:eu-repo/semantics/altIdentifier/doi/10.1071/RD07172
info:eu-repo/semantics/altIdentifier/url/http://www.provino.com.ar/reproduccion-r/366-a-unique-method-to-produce-transgenic-embryos-in-ovine-porcine-feline-bovine-and-equine-species
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Csiro Publishing
publisher.none.fl_str_mv Csiro Publishing
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.070432

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