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Toehold integrated molecular beacon system for a versatile non-enzymatic application

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Abstract

A molecular beacon (MB) is an oligonucleotide hybridization probe with a hairpin-shaped structure that leads to specific and instantaneous nucleic acid hybridization, enabling a variety of applications. However, integration of additional module sequences interferes with the performance of MBs and increases the complexity of sequence design. Herein, we develop and characterize a toehold integrated molecular beacon (ToMB) strategy for nucleic acid hybridization, where the reaction rate can be flexibly regulated by a target-induced MB conformational switch. Using this basic mechanism, the ToMB is capable of identifying nucleic acids with high specificity and a wider linearity range compared with the conventional molecular beacon system. We further applied the ToMB to the construction of a hybridization chain reaction system and a basic OR logic gate VJHto explore its programmability and versatility. Our results strongly suggest that the novel ToMB can act as a powerful nano-module to construct universal and multifunctional biosensors or molecular computations.

Molecular beacon is employed as a flexible and switchable spacer to control the toehold-mediated strand displacement reaction.

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Acknowledgments

We thank the Clinical Research Center from the Second Affiliated Hospital of Zhejiang University School of Medicine for essential technical supports. We thank the American Journal Experts (AJE) for English language editing.

Funding

This study was supported by grant from Natural Science Foundation of Zhejiang Province (Grant nos. LY14H200002, LY15H200002, and LY16H160023).

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Author notes

  1. Zhenping Liu, Yiyun Wang and Xuchu Wang contributed equally to this work.

Authors and Affiliations

  1. Zhejiang Provincial Key Laboratory of Medical Genetics, Key Laboratory of Laboratory Medicine, Ministry of Education, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, Zhejiang, 325035, China

    Zhenping Liu, Yiyun Wang & Zhihua Tao

  2. Department of Laboratory Medicine, the Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, China

    Xuchu Wang, Weiwei Liu, Yibei Dai, Pan Yu, Ying Ping & Zhihua Tao

  3. Department of Transfusion, the Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, China

    Zhaoping Liao

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  1. Zhenping Liu
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  2. Yiyun Wang
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  4. Weiwei Liu
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  9. Zhihua Tao
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Correspondence to Zhihua Tao.

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Liu, Z., Wang, Y., Wang, X. et al. Toehold integrated molecular beacon system for a versatile non-enzymatic application. Anal Bioanal Chem 410, 7285–7293 (2018). https://doi.org/10.1007/s00216-018-1340-z

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  • DOI: https://doi.org/10.1007/s00216-018-1340-z

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