Exp Mol Med.
2014 May;46(5):e93. 10.1038/emm.2014.14.
Simultaneous deletion of floxed genes mediated by CaMKIIalpha-Cre in the brain and in male germ cells: application to conditional and conventional disruption of Goalpha
- Affiliations
-
- 1Departments of Anatomy, Ajou University, School of Medicine, Suwon, South Korea. hysuh@ajou.ac.kr
- 2Neuroscience Graduate Program, Ajou University, School of Medicine, Suwon, South Korea.
- 3BK21-Division of Cell Transformation and Restoration, Ajou University, School of Medicine, Suwon, South Korea.
- 4Center for Cell Death Regulating Biodrug, Ajou University, School of Medicine, Suwon, South Korea.
- 5Department of Molecular Science and Technology, Ajou University, Suwon, South Korea.
- 6National Institute of Environmental Health Sciences, National Institutes of Health, Department of Health and Human Services, Research Triangle Park, NC, USA.
- KMID: 2155358
- DOI: http://doi.org/10.1038/emm.2014.14
Abstract
- 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 (CaMKIIalpha) 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 CaMKIIalpha-Cre can be utilized for simultaneous inactivation of genes in the adult brain and in male germ cells. Double transgenic Rosa26(+/stop-lacZ)::CaMKIIalpha-Cre(+/Cre) mice generated by crossing CaMKIIalpha-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)::CaMKIIalpha-Cre(+/Cre) mice. Similarly, when double transgenic Gnao(+/f)::CaMKIIalpha-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(Delta)) without inheriting the Cre transgene. The Gnao(Delta/Delta) mice closely resembled conventional Go-alpha knockout mice (Gnao(-/-)) with respect to impairment of their behavior. Thus, we conclude that CaMKIIalpha-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 CaMKIIalpha-Cre mice as breeding pairs.
Keyword
- brain; Cre; CaMKII alpha; Gnao; testis
MeSH Terms
-
Animals
Brain/*metabolism
Calcium-Calmodulin-Dependent Protein Kinase Type 2/genetics
Female
GTP-Binding Protein alpha Subunits, Gi-Go/*genetics
*Gene Deletion
Gene Knockout Techniques/*methods
Male
Mice
RNA, Untranslated/genetics
Recombination, Genetic
Spermatozoa/*metabolism
Calcium-Calmodulin-Dependent Protein Kinase Type 2
GTP-Binding Protein alpha Subunits, Gi-Go
RNA, Untranslated
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