A multiplexed, electrochemical interface for gene-circuit-based sensors.

A multiplexed, electrochemical interface for gene-circuit-based sensors.
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DOI:
10.1038/s41557-019-0366-y
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发表时间:
2020-01
期刊:
影响因子:
21.8
通讯作者:
Pardee K
Pardee K
中科院分区:
化学1区
文献类型:
--
作者:
Sadat Mousavi P;Smith SJ;Chen JB;Karlikow M;Tinafar A;Robinson C;Liu W;Ma D;Green AA;Kelley SO;Pardee K

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合成生物学领域已经使用合成基因网络的工程组装来在生物系统中产生广泛的功能。作为这项工作的一部分,基于基因电路的传感器主要使用光学蛋白(例如荧光,比色)作为报告输出,这限制了测量多个不同信号的潜力。在这里,我们提出了一个电化学接口,允许扩展的无细胞基因电路为基础的传感器的多路报告。我们已经设计了一个可扩展的报告酶系统,该系统在溶液中切割特定的DNA序列,当这些新释放的链被捕获在纳米结构的微电极表面时,会产生电化学信号。我们描述了这个接口的发展,并显示其效用使用配体诱导基因电路和立足点开关为基础的传感器,包括检测多个抗生素耐药基因并行。这项技术有可能通过提供材料,硬件和软件的接口来扩展合成生物学领域。
The field of synthetic biology has used the engineered assembly of synthetic gene networks to create a wide range of function in biological systems. As part of this work, gene circuit-based sensors have primarily used optical proteins (e.g. fluorescent, colorimetric) as reporter outputs, which has limited the potential to measure multiple distinct signals. Here we present an electrochemical interface that permits expanded multiplexed reporting for cell-free gene circuit-based sensors. We have engineered a scalable system of reporter enzymes that cleave specific DNA sequences in solution, which results in an electrochemical signal when these newly liberated strands are captured at the surface of a nanostructured microelectrode. We describe the development of this interface and show its utility using a ligand-inducible gene circuit and toehold switch-based sensors, including the detection of multiple antibiotic resistance genes in parallel. This technology has the potential to expand the field of synthetic biology by providing an interface with materials, hardware and software.
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