Issue 3, 2018

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

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

Supplementary files

Article information

Article type
Edge Article
Submitted
21 Jul 2017
Accepted
18 Oct 2017
First published
27 Nov 2017
This article is Open Access

All publication charges for this article have been paid for by the Royal Society of Chemistry
Creative Commons BY-NC license

Chem. Sci., 2018,9, 760-769

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, 9, 760 DOI: 10.1039/C7SC03190D

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