Targeted JAM-C deletion in germ cells by Spo11-controlled Cre recombinase
Targeted JAM-C deletion in germ cells by Spo11-controlled Cre recombinase
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DOI：
10.1242/jcs.072959
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发表时间：
2011-01-01
影响因子：
4
通讯作者：
Nussenzweig, Andre
中科院分区：
文献类型：
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作者：
Pellegrini, Manuela;Claps, Giuseppina;Nussenzweig, Andre
减数分裂是功能配子产生的关键过程。然而，许多基因在减数分裂阶段表达的生物学意义仍然知之甚少，主要是因为基因敲除小鼠的致死表型。由于缺乏合适的Cre转基因株系，使用条件基因敲除方法对这类基因的功能分析受到阻碍。在此，我们描述了在减数分裂Spo11基因控制下表达Cre重组酶的转基因小鼠的产生。利用LacZ-R26(LoxP)和EYFP-R26(LoxP)报告小鼠，我们展示了Cre在雄性和雌性减数分裂过程中的特异性表达和活性。然后，将Spo11(Cre)小鼠与Nbs1和Jam-C小鼠杂交，产生条件性基因敲除。睾丸组织中Nbs1和JAM-C蛋白水平显著降低。虽然Nbs1基因缺失的小鼠出现了轻微的性腺异常，但JAM-C基因敲除小鼠表现出精子生成停滞，正如之前对空白小鼠的描述。这些结果提供了强有力的证据表明，Spo11(Cre)转基因小鼠是一种强大的工具，可以在减数分裂和/或减数分裂后生殖细胞中删除感兴趣的基因。
Meiosis is a crucial process for the production of functional gametes. However, the biological significance of many genes expressed during the meiotic phase remains poorly understood, mainly because of the lethal phenotypes of the knockout mice. Functional analysis of such genes using the conditional knockout approach is hindered by the lack of suitable Cre transgenic lines. We describe here the generation of transgenic mice expressing Cre recombinase under the control of the meiotic Spo11 gene. Using LacZ-R26(loxP) and EYFP-R26(loxP) reporter mice, we show the specific expression and activity of Cre during meiosis in males and females. Spo11(Cre) mice were then crossed with floxed Nbs1 and JAM-C mice to produce conditional knockouts. A strong reduction of Nbs1 and JAM-C protein levels was found in the testis. Although Nbs1-deleted mice developed minor gonadal abnormalities, JAM-C-knockout mice showed a spermiogenetic arrest, as previously described for the null mice. These results provide strong evidence that Spo11(Cre) transgenic mice represent a powerful tool for deleting genes of interest specifically in meiotic and/or in postmeiotic germ cells.