DNA Curtains Shed Light on Complex Molecular Systems During Homologous Recombination
DNA Curtains Shed Light on Complex Molecular Systems During Homologous Recombination
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DOI：
10.3791/61320
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发表时间：
2020-06-01
影响因子：
1.2
通讯作者：
Greene, Eric C.
中科院分区：
文献类型：
--
作者：
Meir, Aviv;Kong, Muwen;Greene, Eric C.
同源重组(HR)对于修复所有生物体中的双链DNA断裂(DSB)和停滞的复制叉是重要的。HR的缺陷与人类细胞基因组完整性的丧失和致癌转化密切相关。HR涉及一组复杂的蛋白质的协调作用，其中许多仍然知之甚少。这里描述的这项研究的关键方面是一种名为“DNA窗帘”的技术，这种技术允许在微流控样品室的表面组装排列好的DNA分子。然后，它们可以通过全内反射荧光显微镜(TIRFM)进行可视化。DNA幕是由我们的实验室首创的，它允许直接访问毫秒时间尺度和纳米尺度的时空信息，这是其他方法无法轻易揭示的。DNA窗帘的一个主要优势是它简化了单分子实验中统计相关数据的收集。这项研究继续为细胞如何调节和维护基因组完整性提供新的见解。
Homologous recombination (HR) is important for the repair of double-stranded DNA breaks (DSBs) and stalled replication forks in all organisms. Defects in HR are closely associated with a loss of genome integrity and oncogenic transformation in human cells. HR involves coordinated actions of a complex set of proteins, many of which remain poorly understood. The key aspect of the research described here is a technology called "DNA curtains", a technique which allows for the assembly of aligned DNA molecules on the surface of a microfluidic sample chamber. They can then be visualized by total internal reflection fluorescence microscopy (TIRFM). DNA curtains was pioneered by our laboratory and allows for direct access to spatiotemporal information at millisecond time scales and nanometer scale resolution, which cannot be easily revealed through other methodologies. A major advantage of DNA curtains is that it simplifies the collection of statistically relevant data from single molecule experiments. This research continues to yield new insights into how cells regulate and preserve genome integrity.