A Strategy Employing a TF-Splinting Duplex Nanoswitch to Achieve Single-Step, Enzyme-Free, Signal-On Detection of l-Tryptophan.

Xuying Duan, Ziwei Chen, Suming Tang, Meiqiong Ge, Hua Wei, Yifu Guan, Guojie Zhao

ACS sensors 2020 Mar 27

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Transcription factor (TF)-based metabolite detection mainly depends on TF-regulated gene expression in cells. From TF activation to gene transcription and translation, the signal travels a relatively long way before it is received. Here, we propose a TF-splinting duplex DNA nanoswitch to detect metabolites. We show its feasibility using tryptophan repressor (TrpR) to detect l-tryptophan as a model. The assay has been optimized and characterized after obtaining a proof of concept, and the detection of l-tryptophan in complex biological samples is feasible. Unlike an equivalent gene expression approach, the whole process is a single-step, enzyme-free, and signal-on method. It can be completed within 20 min. This proposed TF-splinting duplex has the potential to be applied to the quick and convenient detection of other metabolites or even TFs.

Citation

Xuying Duan, Ziwei Chen, Suming Tang, Meiqiong Ge, Hua Wei, Yifu Guan, Guojie Zhao. A Strategy Employing a TF-Splinting Duplex Nanoswitch to Achieve Single-Step, Enzyme-Free, Signal-On Detection of l-Tryptophan. ACS sensors. 2020 Mar 27;5(3):837-844