KMID : 0620920140460050001
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Experimental & Molecular Medicine 2014 Volume.46 No. 5 p.1 ~ p.0
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Simultaneous deletion of floxed genes mediated by CaMKII¥á-Cre in the brain and in male germ cells: application to conditional and conventional disruption of Go¥á
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Choi Chan-Il
Yoon Sang-Phil Choi Jung-Mi Kim Sung-Soo Lee Young-Don Lutz Birnbaumer Suh-Kim Haeyoung
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Abstract
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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 (CaMKII¥á) 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 CaMKII¥á-Cre can be utilized for simultaneous inactivation of genes in the adult brain and in male germ cells. Double transgenic Rosa26+/stop-lacZ::CaMKII¥á-Cre+/Cre mice generated by crossing CaMKII¥á-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::CaMKII¥á-Cre+/Cre mice. Similarly, when double transgenic Gnao+/f::CaMKII¥á-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 (Gnao¥Ä) without inheriting the Cre transgene. The Gnao¥Ä/¥Ä mice closely resembled conventional Go-alpha knockout mice (Gnao?/?) with respect to impairment of their behavior. Thus, we conclude that CaMKII¥á-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 CaMKII¥á-Cre mice as breeding pairs.
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KEYWORD
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brain, Cre, CaMKII alpha, Gnao, testis
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