【2h】

A mitotic recombination system for mouse chromosome 17

机译:小鼠17号染色​​体的有丝分裂重组系统

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摘要

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 ≈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.
机译:同源染色体之间的有丝分裂重组是用于模型生物中镶嵌分析的遗传技术。该技术在小鼠中的普遍应用取决于对单个染色体的有效重组系统的建立以及检测重组事件的可靠而灵敏的方法。在这里,我们在小鼠中建立了Cre / LoxP介导的重组系统,用于全长染色体17的镶嵌分析。在后代测试中,同源染色体之间的Cre介导的种系重组以≈9%的频率观察到。通过表面标记和荧光激活的细胞分选,评估了体细胞组织中的有丝分裂重组,并在B和T淋巴细胞中对其进行了评分。我们表明,沿袭特异性的Cre可以诱导有丝分裂重组,在淋巴样祖细胞中的重现频率为0.5–1.0%。此处建立的重组系统可以简单,准确地检测和分离活细胞中的重组事件,从而使该系统特别适合免疫系统中的镶嵌分析或诱变研究。

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