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.
- Author:
Chan Il CHOI
1
;
Sang Phil YOON
;
Jung Mi CHOI
;
Sung Soo KIM
;
Young Don LEE
;
Lutz BIRNBAUMER
;
Haeyoung SUH-KIM
Author Information
- Publication Type:Original Article ; Research Support, N.I.H., Intramural ; Research Support, Non-U.S. Gov't
- Keywords: brain; Cre; CaMKII alpha; Gnao; testis
- MeSH: 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
- From:Experimental & Molecular Medicine 2014;46(5):e93-
- CountryRepublic of Korea
- Language:English
- 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.
