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Exonuclease III-propelled DNAzyme walker: an electrochemical strategy for microRNA diagnostics

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Abstract

MicroRNA detection is crucial for early infectious disease diagnosis and rapid cancer screening. However, conventional techniques like reverse transcription-quantitative polymerase chain reaction, requiring specialized training and intricate procedures, are less suitable for point-of-care analyses. To address this, we’ve developed a straightforward amplifier based on an exonuclease III (exo III)-propelled DNAzyme walker for sensitive and selective microRNA detection. This amplifier employs a specially designed hairpin probe with two exposed segments for strand recognition. Once the target microRNA is identified by the hairpin’s extended single-strand DNA, exo III initiates its digestion, allowing microRNA regeneration and subsequent hairpin probe digestion cycles. This cyclical process produces a significant amount of DNAzyme, leading to a marked reduction in electrochemical signals. The biosensor exhibits a detection range from 10 fM to 100 pM and achieves a detection limit of 5 fM (3σ criterion). Importantly, by integrating an “And logic gate,” our system gains the capacity for simultaneous diagnosis of multiple microRNAs, enhancing its applicability in RNA-based disease diagnostics.

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Funding

This work is financially supported the Natural Science Foundation of China (51572044, 21475018), and the Fundamental Research Fund for the Central Universities (N160504010).

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Authors and Affiliations

  1. Department of Rehabilitation Medicine, the Affiliated Huaian No.1 People’s Hospital of Nan**g Medical University, Huaian, 223300, China

    Dengke Li

  2. Department of Chemistry, New York University, New York, NY, 10003, USA

    Qiuyan Huang

  3. Department of Physics, New York University, New York, NY, 10003, USA

    Kun Wang

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  1. Dengke Li
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  2. Qiuyan Huang
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Correspondence to Dengke Li.

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Li, D., Huang, Q. & Wang, K. Exonuclease III-propelled DNAzyme walker: an electrochemical strategy for microRNA diagnostics. Microchim Acta 191, 173 (2024). https://doi.org/10.1007/s00604-024-06208-4

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