Optimization of transgenesis conditions for the generation of CXCL2-luciferase reporter mice line

Autores
Crispo, Martina; Schlapp, Geraldine; Cárdenas Rodriguez, Magdalena; González Maciel, María Dolores; Rumbo, Martín
Año de publicación
2013
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Background: Transgenesis by microinjection has been widely used for the generation of different mouse models. Different variables of the procedure may critically affect the efficiency of the process. A DNA construction that carries the CXCL2 promoter gene and firefly luciferase has been used to optimize aspects of the procedure. Three different concentrations (0.5, 1.0 and 4.0 ng/µl) of the DNA construction to microinject a total of 1981 zygotes has been tested. Intact/injected embryos, pregnancy and birth rate, survival of pups 7 days after birth, number of transgenic pups and overall transgenic efficiency was registered and analyzed by Z test of proportions for each group. Results: A total of seven transgenic founders were detected for the three DNA concentrations used, 1 in 46 alive pups in the 0.5 ng/µl group, 5 in 38 alive pups in the 1 ng/µl group and 1 in 21 alive pups in the 4 ng/µl group ( p < 0.1). The overall transgenic efficiency was higher for the 1 ng/µl concentration, with a transgenic rate of 13.2%. Conclusions: In conclusion, we have selected the best operative conditions to maximize the transgenesis efficiency. Furthermore, the transgenic lines developed could be used as a reporter model of innate immunity activation with many different applications in the fields of immunology, cancer and neurodegenerative diseases.
Fil: Crispo, Martina. Instituto Pasteur de Montevideo; Uruguay
Fil: Schlapp, Geraldine. Instituto Pasteur de Montevideo; Uruguay
Fil: Cárdenas Rodriguez, Magdalena. Instituto Pasteur de Montevideo; Uruguay
Fil: González Maciel, María Dolores. Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biologicas. Laboratorio de Investigaciones del Sistema Inmune; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Rumbo, Martín. Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biologicas. Laboratorio de Investigaciones del Sistema Inmune; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Materia
DNA concentration
Microinjection
Reporter systems
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/23540
Acceder
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network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Optimization of transgenesis conditions for the generation of CXCL2-luciferase reporter mice lineCrispo, MartinaSchlapp, GeraldineCárdenas Rodriguez, MagdalenaGonzález Maciel, María DoloresRumbo, MartínDNA concentrationMicroinjectionReporter systemshttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Background: Transgenesis by microinjection has been widely used for the generation of different mouse models. Different variables of the procedure may critically affect the efficiency of the process. A DNA construction that carries the CXCL2 promoter gene and firefly luciferase has been used to optimize aspects of the procedure. Three different concentrations (0.5, 1.0 and 4.0 ng/µl) of the DNA construction to microinject a total of 1981 zygotes has been tested. Intact/injected embryos, pregnancy and birth rate, survival of pups 7 days after birth, number of transgenic pups and overall transgenic efficiency was registered and analyzed by Z test of proportions for each group. Results: A total of seven transgenic founders were detected for the three DNA concentrations used, 1 in 46 alive pups in the 0.5 ng/µl group, 5 in 38 alive pups in the 1 ng/µl group and 1 in 21 alive pups in the 4 ng/µl group ( p < 0.1). The overall transgenic efficiency was higher for the 1 ng/µl concentration, with a transgenic rate of 13.2%. Conclusions: In conclusion, we have selected the best operative conditions to maximize the transgenesis efficiency. Furthermore, the transgenic lines developed could be used as a reporter model of innate immunity activation with many different applications in the fields of immunology, cancer and neurodegenerative diseases.Fil: Crispo, Martina. Instituto Pasteur de Montevideo; UruguayFil: Schlapp, Geraldine. Instituto Pasteur de Montevideo; UruguayFil: Cárdenas Rodriguez, Magdalena. Instituto Pasteur de Montevideo; UruguayFil: González Maciel, María Dolores. Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biologicas. Laboratorio de Investigaciones del Sistema Inmune; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Rumbo, Martín. Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biologicas. Laboratorio de Investigaciones del Sistema Inmune; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaPontificia Universidad Católica de Valparaíso2013-07info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/23540Crispo, Martina; Schlapp, Geraldine; Cárdenas Rodriguez, Magdalena; González Maciel, María Dolores; Rumbo, Martín; Optimization of transgenesis conditions for the generation of CXCL2-luciferase reporter mice line; Pontificia Universidad Católica de Valparaíso; Electronic Journal of Biotechnology; 16; 6; 7-2013; 1-80717-3458CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.2225/vol16-issue6-fulltext-3info:eu-repo/semantics/altIdentifier/url/http://www.ejbiotechnology.info/index.php/ejbiotechnology/article/view/1288info:eu-repo/semantics/altIdentifier/url/http://ref.scielo.org/69hzdbinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:29:30Zoai:ri.conicet.gov.ar:11336/23540instacron: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 10:29:30.874CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Optimization of transgenesis conditions for the generation of CXCL2-luciferase reporter mice line
title Optimization of transgenesis conditions for the generation of CXCL2-luciferase reporter mice line
spellingShingle Optimization of transgenesis conditions for the generation of CXCL2-luciferase reporter mice line
Crispo, Martina
DNA concentration
Microinjection
Reporter systems
title_short Optimization of transgenesis conditions for the generation of CXCL2-luciferase reporter mice line
title_full Optimization of transgenesis conditions for the generation of CXCL2-luciferase reporter mice line
title_fullStr Optimization of transgenesis conditions for the generation of CXCL2-luciferase reporter mice line
title_full_unstemmed Optimization of transgenesis conditions for the generation of CXCL2-luciferase reporter mice line
title_sort Optimization of transgenesis conditions for the generation of CXCL2-luciferase reporter mice line
dc.creator.none.fl_str_mv Crispo, Martina
Schlapp, Geraldine
Cárdenas Rodriguez, Magdalena
González Maciel, María Dolores
Rumbo, Martín
author Crispo, Martina
author_facet Crispo, Martina
Schlapp, Geraldine
Cárdenas Rodriguez, Magdalena
González Maciel, María Dolores
Rumbo, Martín
author_role author
author2 Schlapp, Geraldine
Cárdenas Rodriguez, Magdalena
González Maciel, María Dolores
Rumbo, Martín
author2_role author
author
author
author
dc.subject.none.fl_str_mv DNA concentration
Microinjection
Reporter systems
topic DNA concentration
Microinjection
Reporter systems
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Background: Transgenesis by microinjection has been widely used for the generation of different mouse models. Different variables of the procedure may critically affect the efficiency of the process. A DNA construction that carries the CXCL2 promoter gene and firefly luciferase has been used to optimize aspects of the procedure. Three different concentrations (0.5, 1.0 and 4.0 ng/µl) of the DNA construction to microinject a total of 1981 zygotes has been tested. Intact/injected embryos, pregnancy and birth rate, survival of pups 7 days after birth, number of transgenic pups and overall transgenic efficiency was registered and analyzed by Z test of proportions for each group. Results: A total of seven transgenic founders were detected for the three DNA concentrations used, 1 in 46 alive pups in the 0.5 ng/µl group, 5 in 38 alive pups in the 1 ng/µl group and 1 in 21 alive pups in the 4 ng/µl group ( p < 0.1). The overall transgenic efficiency was higher for the 1 ng/µl concentration, with a transgenic rate of 13.2%. Conclusions: In conclusion, we have selected the best operative conditions to maximize the transgenesis efficiency. Furthermore, the transgenic lines developed could be used as a reporter model of innate immunity activation with many different applications in the fields of immunology, cancer and neurodegenerative diseases.
Fil: Crispo, Martina. Instituto Pasteur de Montevideo; Uruguay
Fil: Schlapp, Geraldine. Instituto Pasteur de Montevideo; Uruguay
Fil: Cárdenas Rodriguez, Magdalena. Instituto Pasteur de Montevideo; Uruguay
Fil: González Maciel, María Dolores. Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biologicas. Laboratorio de Investigaciones del Sistema Inmune; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Rumbo, Martín. Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biologicas. Laboratorio de Investigaciones del Sistema Inmune; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
description Background: Transgenesis by microinjection has been widely used for the generation of different mouse models. Different variables of the procedure may critically affect the efficiency of the process. A DNA construction that carries the CXCL2 promoter gene and firefly luciferase has been used to optimize aspects of the procedure. Three different concentrations (0.5, 1.0 and 4.0 ng/µl) of the DNA construction to microinject a total of 1981 zygotes has been tested. Intact/injected embryos, pregnancy and birth rate, survival of pups 7 days after birth, number of transgenic pups and overall transgenic efficiency was registered and analyzed by Z test of proportions for each group. Results: A total of seven transgenic founders were detected for the three DNA concentrations used, 1 in 46 alive pups in the 0.5 ng/µl group, 5 in 38 alive pups in the 1 ng/µl group and 1 in 21 alive pups in the 4 ng/µl group ( p < 0.1). The overall transgenic efficiency was higher for the 1 ng/µl concentration, with a transgenic rate of 13.2%. Conclusions: In conclusion, we have selected the best operative conditions to maximize the transgenesis efficiency. Furthermore, the transgenic lines developed could be used as a reporter model of innate immunity activation with many different applications in the fields of immunology, cancer and neurodegenerative diseases.
publishDate 2013
dc.date.none.fl_str_mv 2013-07
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/11336/23540
Crispo, Martina; Schlapp, Geraldine; Cárdenas Rodriguez, Magdalena; González Maciel, María Dolores; Rumbo, Martín; Optimization of transgenesis conditions for the generation of CXCL2-luciferase reporter mice line; Pontificia Universidad Católica de Valparaíso; Electronic Journal of Biotechnology; 16; 6; 7-2013; 1-8
0717-3458
CONICET Digital
CONICET
url http://hdl.handle.net/11336/23540
identifier_str_mv Crispo, Martina; Schlapp, Geraldine; Cárdenas Rodriguez, Magdalena; González Maciel, María Dolores; Rumbo, Martín; Optimization of transgenesis conditions for the generation of CXCL2-luciferase reporter mice line; Pontificia Universidad Católica de Valparaíso; Electronic Journal of Biotechnology; 16; 6; 7-2013; 1-8
0717-3458
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.2225/vol16-issue6-fulltext-3
info:eu-repo/semantics/altIdentifier/url/http://www.ejbiotechnology.info/index.php/ejbiotechnology/article/view/1288
info:eu-repo/semantics/altIdentifier/url/http://ref.scielo.org/69hzdb
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Pontificia Universidad Católica de Valparaíso
publisher.none.fl_str_mv Pontificia Universidad Católica de Valparaíso
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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