Despite having many key roles in cellular biology, directly imaging biologically important RNAs has been hindered by a lack of fluorescent tools equivalent to the fluorescent proteins available to study cellular proteins. Ideal RNA labelling systems must preserve biological function, have photophysical properties similar to existing fluorescent proteins, and be compatible with established live and fixed cell protein labelling strategies. Here, we report a microfluidics-based selection of three new high-affinity RNA Mango fluorogenic aptamers. Two of these are as bright or brighter than enhanced GFP when bound to TO1-Biotin. Furthermore, we show that the new Mangos can accurately image the subcellular localization of three small non-coding RNAs (5S, U6, and a box C/D scaRNA) in fixed and live mammalian cells. These new aptamers have many potential applications to study RNA function and dynamics both in vitro and in mammalian cells.
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机译：尽管在细胞生物学中起着许多关键作用，但是由于缺乏与研究细胞蛋白可用的荧光蛋白相当的荧光工具，直接成像生物学上重要的RNA受到了阻碍。理想的RNA标记系统必须保留生物学功能，具有与现有荧光蛋白相似的光物理特性，并与已建立的活细胞和固定细胞蛋白标记策略兼容。在这里，我们报告基于微流控技术的三个新的高亲和力RNA芒果荧光适体的选择。当与TO1-生物素结合时，其中两个与增强的GFP一样明亮。此外，我们表明，新的芒果可以在固定和活的哺乳动物细胞中准确成像三个小的非编码RNA（5S，U6和盒C / D scaRNA）的亚细胞定位。这些新的适体在研究体外和哺乳动物细胞中的RNA功能和动力学方面具有许多潜在的应用。
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