Fluorescence facilitates the detection, visualization, and tracking of molecules with high sensitivity and specificity. A functional DNA molecule that generates a robust fluorescent signal would offer significant advantages for many applications compared to intrinsically fluorescent proteins, which are expensive and labor intensive to synthesize, and fluorescent RNA aptamers, which are unstable under most conditions. Here, we describe a novel deoxyriboyzme that rapidly and efficiently generates a stable fluorescent product using a readily available coumarin substrate. An engineered version can detect picomolar concentrations of ribonucleases in a simple homogenous assay, and was used to rapidly identify novel inhibitors of the SARS-CoV-2 ribonuclease Nsp15 in a high-throughput screen. Our work adds an important new component to the toolkit of functional DNA parts, and also demonstrates how catalytic DNA motifs can be used to solve real-world problems.

中文翻译：

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Aurora：一种用于高通量筛选的荧光脱氧核酶


荧光有助于以高灵敏度和特异性检测、可视化和跟踪分子。与合成昂贵且劳动强度大的内在荧光蛋白和在大多数条件下不稳定的荧光RNA适体相比，产生强大荧光信号的功能性DNA分子将为许多应用提供显着的优势。在这里，我们描述了一种新型脱氧核酶，它使用易于获得的香豆素底物快速有效地产生稳定的荧光产物。工程版本可以通过简单的均质测定检测皮摩尔浓度的核糖核酸酶，并用于在高通量筛选中快速识别 SARS-CoV-2 核糖核酸酶 Nsp15 的新型抑制剂。我们的工作为功能性 DNA 部分的工具箱添加了一个重要的新组件，并且还展示了如何使用催化 DNA 基序来解决现实世界的问题。