Incomplete Cre-mediated excision leads to phenotypic differences between Stra8-iCre; Mov10l1lox/lox and Stra8-iCre; Mov10l1lox/Δ mice

摘要

In the Cre-loxp system, expression level and activity of Cre recombinase in a Cre deletor line are critical because these determine not only the cell specificity of gene knockout (KO), but also the efficiency of Cre-mediated excision in a specific cell lineage. Although the spatiotemporal expression pattern of a Cre transgene is usually defined upon the generation of the mouse line, the Cre excision efficiency in a specific targeted cell lineage is rarely evaluated and often assumed to be 100%. Incomplete excision can lead to highly variable phenotypes due to mosaicism (i.e. co-existence of cells with the flox or the recombined flox allele) and this problem has long been overlooked. Here, we report that Stra8-iCre, a transgenic allele expressing codon-improved Cre recombinase (iCre) under the control of the male germ cell-specific Stra8 promoter, could efficiently delete one Mov10l1 flox allele in spermatogenic cells, whereas the excision was incomplete when two Mov10l1 flox alleles were present. The incomplete Cre-mediated excision led to a testicular phenotype that was much less severe than that in the true conditional KO (100% inactivation) mice. Our findings suggest that it is essential to determine the efficiency of Cre excision when Cre-loxp system is used for deleting genes in a specific cell lineage and the Cre; gene^{lox / Δ} genotype should be used to evaluate phenotypes instead of Cre; gene^lox/lox due to the fact that the latter usually bears incomplete deletion of the flox allele(s).