Simultaneous deletion of floxed genes mediated by CaMKIIα-Cre in the brain and in male germ cells: application to conditional and conventional disruption of Goα.
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 (Gnao(f/f)) were backcrossed to wild-type females, approximately 22% of the offspring carried the disrupted allele (Gnao(Δ)) without inheriting the Cre transgene. The Gnao(Δ/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.
1] Departments of Anatomy, Ajou University, School of Medicine, Suwon, South Korea [2] Neuroscience Graduate Program, Ajou University, School of Medicine, Suwon, South Korea [3] BK21-Division of Cell Transformation and Restoration, Ajou University, School of Medicine, Suwon, South Korea.
Chan-Il
CI
Choi
1] Departments of Anatomy, Ajou University, School of Medicine, Suwon, South Korea [2] Neuroscience Graduate Program, Ajou University, School of Medicine, Suwon, South Korea [3] BK21-Division of Cell Transformation and Restoration, Ajou University, School of Medicine, Suwon, South Korea.
Sang-Phil
SP
Yoon
1] Departments of Anatomy, Ajou University, School of Medicine, Suwon, South Korea [2] Neuroscience Graduate Program, Ajou University, School of Medicine, Suwon, South Korea [3] BK21-Division of Cell Transformation and Restoration, Ajou University, School of Medicine, Suwon, South Korea.
Jung-Mi
JM
Choi
1] Departments of Anatomy, Ajou University, School of Medicine, Suwon, South Korea [2] Center for Cell Death Regulating Biodrug, Ajou University, School of Medicine, Suwon, South Korea.
Sung-Soo
SS
Kim
1] Departments of Anatomy, Ajou University, School of Medicine, Suwon, South Korea [2] Center for Cell Death Regulating Biodrug, Ajou University, School of Medicine, Suwon, South Korea [3] Department of Molecular Science and Technology, Ajou University, Suwon, South Korea.
Young-Don
YD
Lee
National Institute of Environmental Health Sciences, National Institutes of Health, Department of Health and Human Services, Research Triangle Park, NC, USA.
Lutz
L
Birnbaumer
1] Departments of Anatomy, Ajou University, School of Medicine, Suwon, South Korea [2] Neuroscience Graduate Program, Ajou University, School of Medicine, Suwon, South Korea [3] BK21-Division of Cell Transformation and Restoration, Ajou University, School of Medicine, Suwon, South Korea.
Haeyoung
H
Suh-Kim
D000818:Animals; D001921:Brain / Q000378:metabolism*; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2 / Q000235:genetics; D005260:Female; D019206:GTP-Binding Protein alpha Subunits, Gi-Go / Q000235:genetics*; D017353:Gene Deletion; D055786:Gene Knockout Techniques / Q000379:methods*; D008297:Male; D051379:Mice; D022661:RNA, Untranslated / Q000235:genetics; D011995:Recombination, Genetic; D013094:Spermatozoa / Q000378:metabolism*
C110853:Gt(ROSA)26Sor non-coding RNA, mouse; D022661:RNA, Untranslated; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D019206:GTP-Binding Protein alpha Subunits, Gi-Go