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Nano Research 2019, 12(5): 1055-1060 https://doi.org/10.1007/s12274-019-2344-5
Research Article | Issue | Published: 07 March 2019

Dual-enzyme-propelled unbounded DNA walking nanomachine for intracellular imaging of lowly expressed microRNA

Show Author's Information Tianshu Chen1,2,§,  Yaoyao Chen1,§,  Huinan Chen1,  Fan Zhang1,  Qianqian Zhang1,  Guifang Chen1,3( ),  Xiaoli Zhu1( )
Center for Molecular Recognition and Biosensing,School of Life Sciences, Shanghai University,Shanghai,200444,China;
Institute of Biomedical Engineering,School of Communication and Information Engineering, Shanghai University,Shanghai,200444,China;
Department of Bioengineering,University of Washington,Seattle, WA,98195,USA;

§ Tianshu Chen and Yaoyao Chen contributed equally to this work.

Keywords:
microRNA, nanoparticles, DNA walker, nanomachine, imaging analysis
Topics:
Biomolecules DNA EnzymesFluorescenceFluorescence microscopyNanoparticles
Cite this article:
Chen T, Chen Y, Chen H, et al. Dual-enzyme-propelled unbounded DNA walking nanomachine for intracellular imaging of lowly expressed microRNA. Nano Research, 2019, 12(5): 1055-1060. https://doi.org/10.1007/s12274-019-2344-5
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Abstract Full text Electronic supplementary material About this article

Despite the progress on the analysis of miRNA either in vitro or in vivo, intracellular imaging of lowly expressed microRNA remains a challenge. Here we develop a novel dual-enzyme-propelled DNA walking nanomachine, which is tailored to accomplish this mission. The nanomachine is constructed with nanoparticles-loaded DNA tracks, on which the targeted miRNA working as a single-foot DNA walker can move autonomously under the catalysis of two cooperative enzymes. Cleavage of the DNA tracks like a "burnt-bridge" mechanism is thereafter triggered, resulting in an amplified fluorescent signal. After the comprehensive study and optimization of the DNA nanomachine, miR-892b, a significantly down-regulated miRNA in breast cancer cells, is selected as a model target. Sensitivity detection in vitro is achieved with a superior detection limit of 4 pM. While being delivered into cells, the DNA nanomachine is available for the imaging of the lowly expressed microRNA, which is totally missing using the conventional fluorescence in situ hybridization (FISH) method. Up-regulation or down-regulation of the miRNA by exogenous regulatory factors can be also well evaluated. This DNA nanomachine provides a competitive approach for the analysis of miRNA, and has the potential to be extended to some other biomolecules.


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About this article

Dual-enzyme-propelled unbounded DNA walking nanomachine for intracellular imaging of lowly expressed microRNA

Show Author's information Tianshu Chen1,2,§, Yaoyao Chen1,§, Huinan Chen1, Fan Zhang1, Qianqian Zhang1, Guifang Chen1,3( ), Xiaoli Zhu1( )
Center for Molecular Recognition and Biosensing,School of Life Sciences, Shanghai University,Shanghai,200444,China;
Institute of Biomedical Engineering,School of Communication and Information Engineering, Shanghai University,Shanghai,200444,China;
Department of Bioengineering,University of Washington,Seattle, WA,98195,USA;

§ Tianshu Chen and Yaoyao Chen contributed equally to this work.

Abstract

Despite the progress on the analysis of miRNA either in vitro or in vivo, intracellular imaging of lowly expressed microRNA remains a challenge. Here we develop a novel dual-enzyme-propelled DNA walking nanomachine, which is tailored to accomplish this mission. The nanomachine is constructed with nanoparticles-loaded DNA tracks, on which the targeted miRNA working as a single-foot DNA walker can move autonomously under the catalysis of two cooperative enzymes. Cleavage of the DNA tracks like a "burnt-bridge" mechanism is thereafter triggered, resulting in an amplified fluorescent signal. After the comprehensive study and optimization of the DNA nanomachine, miR-892b, a significantly down-regulated miRNA in breast cancer cells, is selected as a model target. Sensitivity detection in vitro is achieved with a superior detection limit of 4 pM. While being delivered into cells, the DNA nanomachine is available for the imaging of the lowly expressed microRNA, which is totally missing using the conventional fluorescence in situ hybridization (FISH) method. Up-regulation or down-regulation of the miRNA by exogenous regulatory factors can be also well evaluated. This DNA nanomachine provides a competitive approach for the analysis of miRNA, and has the potential to be extended to some other biomolecules.

Keywords: microRNA, nanoparticles, DNA walker, nanomachine, imaging analysis

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Publication history
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Acknowledgements

Publication history

Received: 21 December 2018
Revised: 17 February 2019
Accepted: 17 February 2019
Published: 07 March 2019
Issue date: May 2019

Copyright

© Tsinghua University Press and Springer-Verlag GmbH Germany, part of Springer Nature 2019

Acknowledgements

Acknowledgements

This work was supported by the National Natural Science Foundation of China (Nos. 81873539 and 21575088) and the Natural Science Foundation of Shanghai (No. 14ZR1416500).

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