Department of Laboratory Medicine and Pathology, Faculty of Medicine and Dentistry, University of Alberta, Edmonton, Alberta, T6G 2G3, Canada; College of Pharmaceutical Sciences, Southwest University, Chongqing, 400715, China.
Department of Laboratory Medicine and Pathology, Faculty of Medicine and Dentistry, University of Alberta, Edmonton, Alberta, T6G 2G3, Canada.
Biosens Bioelectron. 2022 Dec 1;217:114704. doi: 10.1016/j.bios.2022.114704. Epub 2022 Sep 9.
DNAzyme motor systems using gold nanoparticles (AuNPs) as scaffolds are useful for biosensing and in situ amplification because these systems are free of protein enzymes, isothermal, homogeneous, and sensitive. However, detecting different targets using the available DNAzyme motor techniques requires redesigns of the DNAzyme motor. We report here a toehold-exchange translator and the translator-mediated DNAzyme motor systems, which enable sensitive responses to various nucleic acid targets using the same DNAzyme motor without requiring redesign. The translator is able to efficiently convert different nucleic acid targets into a specific output DNA that further activates the pre-silenced DNAzyme motor and consequently initiates the autonomous walking of the DNAzyme motor. Simply adjusting the target-binding region of the translator enables the same DNAzyme motor system to respond to various nucleic acid targets. The translator-mediated DNAzyme motor system is able to detect as low as 2.5 pM microRNA-10b and microRNA-21 under room temperature without the need of separation or washing. We further demonstrate the versatility of the translator and the DNAzyme motor by successful construction and operation of four logic gates, including OR, AND, NOR, and NAND logic gates. These logic gates use two microRNA targets as inputs and generate amplified fluorescence signals from the operation of the same DNAzyme motor. Incorporation of the toehold-exchange translator into the DNAzyme motor technology improves the biosensing applications of DNA motors to diverse nucleic acid targets.
基于金纳米粒子（AuNPs）作为支架的 DNA 酶马达系统在生物传感和原位扩增方面很有用，因为这些系统不含蛋白质酶，具有等温、均相和灵敏的特点。然而，利用现有的 DNA 酶马达技术检测不同的靶标需要重新设计 DNA 酶马达。我们在此报告一种适体交换翻译器和翻译器介导的 DNA 酶马达系统，该系统能够使用相同的 DNA 酶马达对各种核酸靶标进行灵敏响应，而无需重新设计。翻译器能够有效地将不同的核酸靶标转化为特定的输出 DNA，进一步激活预沉默的 DNA 酶马达，并因此启动 DNA 酶马达的自主运动。只需调整翻译器的靶标结合区域，就可以使相同的 DNA 酶马达系统对各种核酸靶标做出响应。翻译器介导的 DNA 酶马达系统能够在室温下检测低至 2.5 pM 的 microRNA-10b 和 microRNA-21，而无需分离或洗涤。我们进一步通过成功构建和操作四个逻辑门，包括 OR、AND、NOR 和 NAND 逻辑门，证明了翻译器和 DNA 酶马达的多功能性。这些逻辑门使用两个 microRNA 靶标作为输入，并从相同的 DNA 酶马达的操作中产生放大的荧光信号。将适体交换翻译器纳入 DNA 酶马达技术提高了 DNA 马达对各种核酸靶标的生物传感应用。
