A mitotic recombination system for mouse chromosome 17
A mitotic recombination system for mouse chromosome 17
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DOI：
10.1073/pnas.0800798105
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发表时间：
2008-03-18
影响因子：
11.1
通讯作者：
Zhuang, Yuan
中科院分区：
文献类型：
--
作者：
Sun, Lei;Wu, Xiaohui;Zhuang, Yuan
同源染色体间的有丝分裂重组是一种用于模式生物嵌合体分析的遗传技术。该技术在小鼠中的普遍应用取决于单个染色体的有效重组系统的建立和用于检测重组事件的可靠且灵敏的方法。在这里，我们建立了一个Cre/LoxP介导的重组系统在小鼠全长染色体17嵌合分析。在后代测试中，观察到同源染色体之间的Cre介导的生殖系重组的频率约为9%。借助于表面标记物和荧光激活细胞分选，在B和T淋巴细胞中评价和评分体细胞组织中的有丝分裂重组。我们表明，谱系特异性Cre可以在淋巴祖细胞中以0.5-1.0%的高度可重复频率诱导有丝分裂重组。在此建立的重组系统允许活细胞中重组事件的简单且准确的检测和分离，使得该系统对于免疫系统中的嵌合分析或诱变研究特别有吸引力。
Mitotic recombination between homologous chromosomes is a genetic technique for mosaic analysis in model organisms. The general application of this technique in the mouse depends on establishment of effective recombination systems for individual chromosomes and reliable and sensitive methods for detection of recombination events. Here, we established a Cre/LoxP-mediated recombination system in mice for mosaic analysis of full-length chromosome 17. Cre-mediated germ-line recombination between the homologous chromosomes was observed with approximate to 9% frequency in a progeny test. Mitotic recombination in somatic tissues was evaluated and scored in B and T lymphocytes with the aid of surface markers and fluorescent-activated cell sorting. We show that a lineage-specific Cre can induce mitotic recombination with a highly reproducible frequency of 0.5-1.0% in lymphoid progenitors. The recombination system established here allows for a simple and accurate detection and isolation of recombination events in live cells, making this system particularly attractive for mosaic analysis or mutagenesis studies in the immune system.