Department of Chemical and Biological Engineering, The Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong.
ACS Synth Biol. 2021 Feb 19;10(2):371-378. doi: 10.1021/acssynbio.0c00545. Epub 2021 Jan 22.
Detection of environmental pollutants is crucial to safeguard ecological and public health. Here, we report a modular biosensing approach for the detection of contaminants based on the regulation of a minimal DNA signal amplifier and transducer circuit by allosteric transcription factors and their cognate ligands. We leverage the competition between allosteric proteins and an endonuclease to modulate cascade toehold-mediated strand displacement reactions, which are triggered in the presence of specific effectors and sustained by the endonuclease. We built two optical biosensors for the detection of tetracyclines and macrolides in water using repressors TetR and MphR, respectively. We demonstrate that our minimal, fast, and single-step biosensors can successfully detect antibiotics in nanomolar levels and apply them to report the presence of spiked-in antibiotics in water samples in a matter of minutes, suggesting great potential for monitoring of water contaminants.
环境污染物的检测对于维护生态和公共健康至关重要。在这里，我们报告了一种基于变构转录因子及其同源配体调节最小 DNA 信号放大器和换能器电路的污染物检测的模块化生物传感方法。我们利用变构蛋白和内切酶之间的竞争来调节级联的链置换反应，该反应在存在特定效应物时被触发，并由内切酶维持。我们使用阻遏物 TetR 和 MphR 分别构建了两种用于检测水中四环素和大环内酯类抗生素的光学生物传感器。我们证明，我们的最小化、快速和单步生物传感器可以成功地检测纳摩尔级别的抗生素，并将其应用于在几分钟内报告水样中添加的抗生素的存在，这表明该方法在监测水中污染物方面具有很大的潜力。
