Adaptive immunity against foreign genetic elements conferred by the CRISPR systems in microbial species has been repurposed as a revolutionary technology for wide-ranging biological applications—chiefly genome engineering. Biochemical, structural, genetic, and genomics studies have revealed important insights into their function and mechanisms, but most ensemble studies cannot observe structural changes of these molecules during their function and are often blind to key reaction intermediates. Here, we review the use of single molecule approaches such as fluorescent particle tracking, FRET, magnetic tweezers, and atomic force microscopy imaging in improving our understanding of the CRISPR toolbox.

中文翻译：

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使用单分子方法了解CRISPR工具箱的分子机制

针对微生物物种中的CRISPR系统赋予的针对外来遗传元素的适应性免疫已被重新定位为一项革命性技术，可广泛应用于生物学应用（主要是基因组工程）。生化，结构，遗传和基因组学研究已经揭示了它们的功能和机制的重要见解，但是大多数集成研究无法观察到这些分子在其功能期间的结构变化，并且通常对关键的反应中间体视而不见。在这里，我们回顾了单分子方法（例如荧光粒子跟踪，FRET，磁性镊子和原子力显微镜成像）在增进我们对CRISPR工具箱的理解中的用途。