Simultaneous deletion of floxed genes mediated by CaMKIIa-Cre in the brain and in male germ cells: application to conditional and conventional disruption of Go-alfa

Autores
Choi, Chan-Il; Yoon, Sang-Phil; Choi, Jung-Mi; Kim, Sung-Soo; Lee, Young-Don; Birnbaumer, Lutz; Suh-Kim. Haeyoung
Año de publicación
2014
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The Cre/LoxP system is a well-established approach to spatially and temporally control genetic inactivation. The calcium/calmodulin-dependent protein kinase II alpha subunit (CaMKIIα) promoter limits expression to specific regions of the forebrain and thus has been utilized for the brain-specific inactivation of the genes. Here, we show that CaMKIIα-Cre can be utilized for simultaneous inactivation of genes in the adult brain and in male germ cells. Double transgenic Rosa26+/stop-lacZ::CaMKIIα-Cre+/Cre mice generated by crossing CaMKIIα-Cre+/Cre mice with floxed ROSA26 lacZ reporter (Rosa26+/stop-lacZ) mice exhibited lacZ expression in the brain and testis. When these mice were mated to wild-type females, about 27% of the offspring were whole body blue by X-gal staining without inheriting the Cre transgene. These results indicate that recombination can occur in the germ cells of male Rosa26+/stop-lacZ::CaMKIIα-Cre+/Cre mice. Similarly, when double transgenic Gnao+/f::CaMKIIα-Cre+/Cre mice carrying a floxed Go-alpha gene (Gnaof/f) were backcrossed to wild-type females, approximately 22% of the offspring carried the disrupted allele (GnaoΔ) without inheriting the Cre transgene. The GnaoΔ/Δ mice closely resembled conventional Go-alpha knockout mice (Gnao−/−) with respect to impairment of their behavior. Thus, we conclude that CaMKIIα-Cre mice afford recombination for both tissue- and time-controlled inactivation of floxed target genes in the brain and for their permanent disruption. This work also emphasizes that extra caution should be exercised in utilizing CaMKIIα-Cre mice as breeding pairs.
Fil: Choi, Chan-Il. Ajou University. School of Medicine; Corea del Sur
Fil: Yoon, Sang-Phil. Ajou University. School of Medicine; Corea del Sur
Fil: Choi, Jung-Mi. Ajou University. School of Medicine; Corea del Sur
Fil: Kim, Sung-Soo. Ajou University. School of Medicine; Corea del Sur
Fil: Lee, Young-Don. Ajou University. School of Medicine; Corea del Sur
Fil: Birnbaumer, Lutz. National Institute of Environmental Health Sciences; Estados Unidos. Pontificia Universidad Católica Argentina "Santa María de los Buenos Aires". Instituto de Investigaciones Biomédicas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas; Argentina
Fil: Suh-Kim. Haeyoung. Ajou University. School of Medicine; Corea del Sur
Materia
BRAIN
CRE
CaMKII ALPHA
GNAO
TESTIS
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:11336/99772
Acceder
id CONICETDig_7ff7ead48a48cf18f3aefe0c88fa7150
oai_identifier_str oai:ri.conicet.gov.ar:11336/99772
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Simultaneous deletion of floxed genes mediated by CaMKIIa-Cre in the brain and in male germ cells: application to conditional and conventional disruption of Go-alfaChoi, Chan-IlYoon, Sang-PhilChoi, Jung-MiKim, Sung-SooLee, Young-DonBirnbaumer, LutzSuh-Kim. HaeyoungBRAINCRECaMKII ALPHAGNAOTESTIShttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The Cre/LoxP system is a well-established approach to spatially and temporally control genetic inactivation. The calcium/calmodulin-dependent protein kinase II alpha subunit (CaMKIIα) promoter limits expression to specific regions of the forebrain and thus has been utilized for the brain-specific inactivation of the genes. Here, we show that CaMKIIα-Cre can be utilized for simultaneous inactivation of genes in the adult brain and in male germ cells. Double transgenic Rosa26+/stop-lacZ::CaMKIIα-Cre+/Cre mice generated by crossing CaMKIIα-Cre+/Cre mice with floxed ROSA26 lacZ reporter (Rosa26+/stop-lacZ) mice exhibited lacZ expression in the brain and testis. When these mice were mated to wild-type females, about 27% of the offspring were whole body blue by X-gal staining without inheriting the Cre transgene. These results indicate that recombination can occur in the germ cells of male Rosa26+/stop-lacZ::CaMKIIα-Cre+/Cre mice. Similarly, when double transgenic Gnao+/f::CaMKIIα-Cre+/Cre mice carrying a floxed Go-alpha gene (Gnaof/f) were backcrossed to wild-type females, approximately 22% of the offspring carried the disrupted allele (GnaoΔ) without inheriting the Cre transgene. The GnaoΔ/Δ mice closely resembled conventional Go-alpha knockout mice (Gnao−/−) with respect to impairment of their behavior. Thus, we conclude that CaMKIIα-Cre mice afford recombination for both tissue- and time-controlled inactivation of floxed target genes in the brain and for their permanent disruption. This work also emphasizes that extra caution should be exercised in utilizing CaMKIIα-Cre mice as breeding pairs.Fil: Choi, Chan-Il. Ajou University. School of Medicine; Corea del SurFil: Yoon, Sang-Phil. Ajou University. School of Medicine; Corea del SurFil: Choi, Jung-Mi. Ajou University. School of Medicine; Corea del SurFil: Kim, Sung-Soo. Ajou University. School of Medicine; Corea del SurFil: Lee, Young-Don. Ajou University. School of Medicine; Corea del SurFil: Birnbaumer, Lutz. National Institute of Environmental Health Sciences; Estados Unidos. Pontificia Universidad Católica Argentina "Santa María de los Buenos Aires". Instituto de Investigaciones Biomédicas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas; ArgentinaFil: Suh-Kim. Haeyoung. Ajou University. School of Medicine; Corea del SurKorean Soc Med Biochemistry Molecular Biology2014-05-02info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/99772Choi, Chan-Il ; Yoon, Sang-Phil; Choi, Jung-Mi; Kim, Sung-Soo; Lee, Young-Don; et al.; Simultaneous deletion of floxed genes mediated by CaMKIIa-Cre in the brain and in male germ cells: application to conditional and conventional disruption of Go-alfa; Korean Soc Med Biochemistry Molecular Biology; Experimental And Molecular Medicine Emm; 46; 2-5-2014; e931226-3613CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.nature.com/articles/emm201414info:eu-repo/semantics/altIdentifier/doi/10.1038/emm.2014.14info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-29T11:41:07Zoai:ri.conicet.gov.ar:11336/99772instacron: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:34982025-10-29 11:41:07.85CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Simultaneous deletion of floxed genes mediated by CaMKIIa-Cre in the brain and in male germ cells: application to conditional and conventional disruption of Go-alfa
title Simultaneous deletion of floxed genes mediated by CaMKIIa-Cre in the brain and in male germ cells: application to conditional and conventional disruption of Go-alfa
spellingShingle Simultaneous deletion of floxed genes mediated by CaMKIIa-Cre in the brain and in male germ cells: application to conditional and conventional disruption of Go-alfa
Choi, Chan-Il
BRAIN
CRE
CaMKII ALPHA
GNAO
TESTIS
title_short Simultaneous deletion of floxed genes mediated by CaMKIIa-Cre in the brain and in male germ cells: application to conditional and conventional disruption of Go-alfa
title_full Simultaneous deletion of floxed genes mediated by CaMKIIa-Cre in the brain and in male germ cells: application to conditional and conventional disruption of Go-alfa
title_fullStr Simultaneous deletion of floxed genes mediated by CaMKIIa-Cre in the brain and in male germ cells: application to conditional and conventional disruption of Go-alfa
title_full_unstemmed Simultaneous deletion of floxed genes mediated by CaMKIIa-Cre in the brain and in male germ cells: application to conditional and conventional disruption of Go-alfa
title_sort Simultaneous deletion of floxed genes mediated by CaMKIIa-Cre in the brain and in male germ cells: application to conditional and conventional disruption of Go-alfa
dc.creator.none.fl_str_mv Choi, Chan-Il
Yoon, Sang-Phil
Choi, Jung-Mi
Kim, Sung-Soo
Lee, Young-Don
Birnbaumer, Lutz
Suh-Kim. Haeyoung
author Choi, Chan-Il
author_facet Choi, Chan-Il
Yoon, Sang-Phil
Choi, Jung-Mi
Kim, Sung-Soo
Lee, Young-Don
Birnbaumer, Lutz
Suh-Kim. Haeyoung
author_role author
author2 Yoon, Sang-Phil
Choi, Jung-Mi
Kim, Sung-Soo
Lee, Young-Don
Birnbaumer, Lutz
Suh-Kim. Haeyoung
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv BRAIN
CRE
CaMKII ALPHA
GNAO
TESTIS
topic BRAIN
CRE
CaMKII ALPHA
GNAO
TESTIS
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv The Cre/LoxP system is a well-established approach to spatially and temporally control genetic inactivation. The calcium/calmodulin-dependent protein kinase II alpha subunit (CaMKIIα) promoter limits expression to specific regions of the forebrain and thus has been utilized for the brain-specific inactivation of the genes. Here, we show that CaMKIIα-Cre can be utilized for simultaneous inactivation of genes in the adult brain and in male germ cells. Double transgenic Rosa26+/stop-lacZ::CaMKIIα-Cre+/Cre mice generated by crossing CaMKIIα-Cre+/Cre mice with floxed ROSA26 lacZ reporter (Rosa26+/stop-lacZ) mice exhibited lacZ expression in the brain and testis. When these mice were mated to wild-type females, about 27% of the offspring were whole body blue by X-gal staining without inheriting the Cre transgene. These results indicate that recombination can occur in the germ cells of male Rosa26+/stop-lacZ::CaMKIIα-Cre+/Cre mice. Similarly, when double transgenic Gnao+/f::CaMKIIα-Cre+/Cre mice carrying a floxed Go-alpha gene (Gnaof/f) were backcrossed to wild-type females, approximately 22% of the offspring carried the disrupted allele (GnaoΔ) without inheriting the Cre transgene. The GnaoΔ/Δ mice closely resembled conventional Go-alpha knockout mice (Gnao−/−) with respect to impairment of their behavior. Thus, we conclude that CaMKIIα-Cre mice afford recombination for both tissue- and time-controlled inactivation of floxed target genes in the brain and for their permanent disruption. This work also emphasizes that extra caution should be exercised in utilizing CaMKIIα-Cre mice as breeding pairs.
Fil: Choi, Chan-Il. Ajou University. School of Medicine; Corea del Sur
Fil: Yoon, Sang-Phil. Ajou University. School of Medicine; Corea del Sur
Fil: Choi, Jung-Mi. Ajou University. School of Medicine; Corea del Sur
Fil: Kim, Sung-Soo. Ajou University. School of Medicine; Corea del Sur
Fil: Lee, Young-Don. Ajou University. School of Medicine; Corea del Sur
Fil: Birnbaumer, Lutz. National Institute of Environmental Health Sciences; Estados Unidos. Pontificia Universidad Católica Argentina "Santa María de los Buenos Aires". Instituto de Investigaciones Biomédicas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas; Argentina
Fil: Suh-Kim. Haeyoung. Ajou University. School of Medicine; Corea del Sur
description The Cre/LoxP system is a well-established approach to spatially and temporally control genetic inactivation. The calcium/calmodulin-dependent protein kinase II alpha subunit (CaMKIIα) promoter limits expression to specific regions of the forebrain and thus has been utilized for the brain-specific inactivation of the genes. Here, we show that CaMKIIα-Cre can be utilized for simultaneous inactivation of genes in the adult brain and in male germ cells. Double transgenic Rosa26+/stop-lacZ::CaMKIIα-Cre+/Cre mice generated by crossing CaMKIIα-Cre+/Cre mice with floxed ROSA26 lacZ reporter (Rosa26+/stop-lacZ) mice exhibited lacZ expression in the brain and testis. When these mice were mated to wild-type females, about 27% of the offspring were whole body blue by X-gal staining without inheriting the Cre transgene. These results indicate that recombination can occur in the germ cells of male Rosa26+/stop-lacZ::CaMKIIα-Cre+/Cre mice. Similarly, when double transgenic Gnao+/f::CaMKIIα-Cre+/Cre mice carrying a floxed Go-alpha gene (Gnaof/f) were backcrossed to wild-type females, approximately 22% of the offspring carried the disrupted allele (GnaoΔ) without inheriting the Cre transgene. The GnaoΔ/Δ mice closely resembled conventional Go-alpha knockout mice (Gnao−/−) with respect to impairment of their behavior. Thus, we conclude that CaMKIIα-Cre mice afford recombination for both tissue- and time-controlled inactivation of floxed target genes in the brain and for their permanent disruption. This work also emphasizes that extra caution should be exercised in utilizing CaMKIIα-Cre mice as breeding pairs.
publishDate 2014
dc.date.none.fl_str_mv 2014-05-02
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/11336/99772
Choi, Chan-Il ; Yoon, Sang-Phil; Choi, Jung-Mi; Kim, Sung-Soo; Lee, Young-Don; et al.; Simultaneous deletion of floxed genes mediated by CaMKIIa-Cre in the brain and in male germ cells: application to conditional and conventional disruption of Go-alfa; Korean Soc Med Biochemistry Molecular Biology; Experimental And Molecular Medicine Emm; 46; 2-5-2014; e93
1226-3613
CONICET Digital
CONICET
url http://hdl.handle.net/11336/99772
identifier_str_mv Choi, Chan-Il ; Yoon, Sang-Phil; Choi, Jung-Mi; Kim, Sung-Soo; Lee, Young-Don; et al.; Simultaneous deletion of floxed genes mediated by CaMKIIa-Cre in the brain and in male germ cells: application to conditional and conventional disruption of Go-alfa; Korean Soc Med Biochemistry Molecular Biology; Experimental And Molecular Medicine Emm; 46; 2-5-2014; e93
1226-3613
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.nature.com/articles/emm201414
info:eu-repo/semantics/altIdentifier/doi/10.1038/emm.2014.14
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
dc.publisher.none.fl_str_mv Korean Soc Med Biochemistry Molecular Biology
publisher.none.fl_str_mv Korean Soc Med Biochemistry Molecular Biology
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
_version_ 1847426302496210944
score 13.10058

Publicaciones similares