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Establishment of a universal and rational gene detection strategy through three-way junction-based remote transduction

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Abstract

The polymerase chain reaction and many isothermal amplifications are able to achieve super gene amplification. Unfortunately, most commonly-used transduction methods, such as dye staining and Taqman-like probing, still suffer from shortcomings including false signals or difficult probe design, or are incompatible with multi-analysis. Here a universal and rational gene detection strategy has been established by translating isothermal amplicons to enzyme-free strand displacement circuits via three-way junction-based remote transduction. An assistant transduction probe was imported to form a partial hybrid with the target single-stranded nucleic acid. After systematic optimization the hybrid could serve as an associative trigger to activate a downstream circuit detector via a strand displacement reaction across the three-way junction. By doing so, the detection selectivity can be double-guaranteed through both amplicon–transducer recognition and the amplicon–circuit reaction. A well-optimized circuit can be immediately applied to a new target detection through simply displacing only 10–12 nt on only one component, according to the target. More importantly, this property for the first time enables multi-analysis and logic-analysis in a single reaction, sharing a single fluorescence reporter. In an applicable model, trace amounts of Cronobacter and Enterobacteria genes have been clearly distinguished from samples with no bacteria or one bacterium, with ultra-high sensitivity and selectivity.

Graphical abstract: Establishment of a universal and rational gene detection strategy through three-way junction-based remote transduction

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Publication details

The article was received on 21 Jul 2017, accepted on 18 Oct 2017 and first published on 27 Nov 2017


Article type: Edge Article
DOI: 10.1039/C7SC03190D
Citation: Chem. Sci., 2018, Advance Article
  • Open access: Creative Commons BY-NC license
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    Establishment of a universal and rational gene detection strategy through three-way junction-based remote transduction

    Y. Tang, B. Lu, Z. Zhu and B. Li, Chem. Sci., 2018, Advance Article , DOI: 10.1039/C7SC03190D

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