Colorimetric assays in which the color of a solution changes in the presence of an input provide a simple and inexpensive way to monitor experimental readouts. In this study we used in vitro selection to identify a self-phosphorylating kinase deoxyribozyme that produces a colorimetric signal by converting the colorless substrate pNPP into the yellow product pNP. The minimized catalytic core, sequence requirements, secondary structure, and buffer requirements of this deoxyribozyme, which we named Apollon, were characterized using a variety of techniques including reselection experiments, high-throughput sequencing, comparative analysis, biochemical activity assays, and NMR. A bimolecular version of Apollon catalyzed multiple turnover phosphorylation and amplified the colorimetric signal. Engineered versions of Apollon could detect oligonucleotides with specific sequences as well as several different types of nucleases in homogenous assays that can be performed in a single tube without the need for washes or purifications. We anticipate that Apollon will be particularly useful to reduce costs in high-throughput screens and for applications in which specialized equipment is not available.

中文翻译：

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Apollon：一种产生黄色产物的脱氧核酶


比色测定中溶液颜色在输入存在时发生变化，提供了一种简单且廉价的方法来监测实验读数。在这项研究中，我们使用体外选择来鉴定一种自磷酸化激酶脱氧核酶，该酶​​通过将无色底物 pNPP 转化为黄色产物 pNP 来产生比色信号。这种脱氧核酶（我们将其命名为 Apollon）的最小化催化核心、序列要求、二级结构和缓冲液要求采用多种技术进行了表征，包括重选择实验、高通量测序、比较分析、生化活性测定和 NMR。 Apollon 的双分子版本催化多次转换磷酸化并放大比色信号。 Apollon 的工程版本可以在同质测定中检测具有特定序列的寡核苷酸以及几种不同类型的核酸酶，这些测定可以在单管中进行，无需洗涤或纯化。我们预计 Apollon 将特别有助于降低高通量筛选的成本以及无法使用专用设备的应用。