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A CRISPR-Cas12a-based electrochemical biosensor for the detection of microphthalmia-associated transcription factor

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A Correction to this article was published on 12 February 2024

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Abstract

A novel electrochemical biosensor that combines the CRISPR-Cas12a system with a gold electrode is reported for the rapid and sensitive detection of microphthalmia-associated transcription factor (MITF). The biosensor consists of a gold electrode modified with DNA1, which contains the target sequence of MITF and is labeled with ferrocene, an electroactive molecule. The biosensor also includes hairpin DNA, which has a binding site for MITF and can hybridize with helper DNA to form a double-stranded complex that activates CRISPR-Cas12a. When MITF is present, it binds to hairpin DNA and prevents its hybridization with helper DNA, thus inhibiting CRISPR-Cas12a activity and preserving the DPV signal of ferrocene. When MITF is absent, hairpin DNA hybridizes with helper DNA and activates CRISPR-Cas12a, which cleaves DNA1 and releases ferrocene, thus reducing the DPV signal. The biosensor can detect MITF with high sensitivity (with an LOD of 8.14 fM), specificity, and accuracy in various samples, such as cell nuclear extracts and human serum. The biosensor can also diagnose and monitor melanocyte-related diseases and melanin production. This work provides a simple, fast, sensitive, and cost-effective biosensor for MITF detection and a valuable tool for applications in genetic testing, disease diagnosis, and drug screening.

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Acknowledgements

We would like to thank the Startup Foundation for Introducing Talent of NUIST (2023r129), the National Natural Science Foundation of China (21964018, 82360259), the Natural Science Foundation of Guangxi Province (2019GXNSFDA245034), and the Guangxi key laboratory of basic and translational research of Bone and joint Degenerative Disease (21-220-06-202205).

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  1. Qisheng Luo and Chunyuan Zhang contributed equally to this work.

Authors and Affiliations

  1. The Affiliated Hospital of Youjiang Medical University for Nationalities, Baise, 533000, Guangxi, China

    Qisheng Luo, Chunyuan Zhang, Xiandong Deng, Dongyuan Liu, Xingchen Pan, Yuanxun Gong & Qianli Tang

  2. School of Chemistry and Materials Science, Nanjing University of Information Science and Technology, Nanjing, 210044, People’s Republic of China

    Kai Zhang

  3. West Guangxi Key Laboratory for Prevention and Treatment of High-Incidence Diseases, Youjiang Medical University for Nationalities, Guangxi, Baise, 533000, China

    Xianjiu Liao

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Correspondence to Qianli Tang, Kai Zhang or Xianjiu Liao.

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The original online version of this article was revised: In this article the affiliation details for Xianjiu Liao were incorrectly given as 'West Guangxi Key Laboratory for Prevention and Treatment of High-Incidence Diseases, Baise 533000, Guangxi, China' but should have been 'West Guangxi Key Laboratory for Prevention and Treatment of High-Incidence Diseases, Youjiang Medical University for Nationalities, Guangxi, Baise, 533000, China'.

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Luo, Q., Zhang, C., Deng, X. et al. A CRISPR-Cas12a-based electrochemical biosensor for the detection of microphthalmia-associated transcription factor. Microchim Acta 191, 73 (2024). https://doi.org/10.1007/s00604-023-06164-5

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