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Issue 26, 2018
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Construction of an enzyme-free concatenated DNA circuit for signal amplification and intracellular imaging

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Abstract

Nucleic acid circuits have shown promising potential for amplified detection of biomarkers with interest in biologically important engineering applications. In this work, by properly integrating two signal amplification approaches, catalytic hairpin assembly (CHA) and hybridization chain reaction (HCR), a concatenated CHA–HCR system was established as an isothermal enzyme-free amplification strategy for highly sensitive and selective nucleic acid assay. The target catalyzes the self-assembly of CHA hairpin substrates into dsDNA products, where the split segments of HCR trigger are successively connected to drive the subsequent autonomous cross-opening of HCR hairpins, leading to the construction of HCR tandem copolymeric dsDNA nanowires. The resulting HCR copolymer brings a fluorophore donor/acceptor pair into close proximity that allows an efficient generation of FRET readout signal. Moreover, the optimized CHA–HCR circuit, upon the incorporation of an auxiliary sensing module, can be converted into a universal sensing platform for detecting cancerous biomarkers (e.g., a well-known oncogene miR-21) through a convenient easy-to-integrate procedure. The concatenated CHA–HCR amplifier enables accurate intracellular miRNA imaging in living cells, which is especially suitable for in situ amplified detection of lowly expressed endogenous analytes. The inherent synergistically accelerated recognition and hybridization features of CHA–HCR circuit contribute to the amplified detection of endogenous RNAs in living cells. The flexible and programmable nature of the homogeneous CHA–HCR system provides a versatile and robust toolbox for a wide range of research fields, such as in vivo bioimaging, clinical diagnosis and environmental monitoring.

Graphical abstract: Construction of an enzyme-free concatenated DNA circuit for signal amplification and intracellular imaging

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Publication details

The article was received on 02 May 2018, accepted on 05 Jun 2018 and first published on 06 Jun 2018


Article type: Edge Article
DOI: 10.1039/C8SC01981A
Citation: Chem. Sci., 2018,9, 5842-5849
  • Open access: Creative Commons BY-NC license
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    Construction of an enzyme-free concatenated DNA circuit for signal amplification and intracellular imaging

    H. Wang, C. Li, X. Liu, X. Zhou and F. Wang, Chem. Sci., 2018, 9, 5842
    DOI: 10.1039/C8SC01981A

    This article is licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported Licence. Material from this article can be used in other publications provided that the correct acknowledgement is given with the reproduced material and it is not used for commercial purposes.

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