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Strand displacement–triggered FRET nanoprobe tracking TK1 mRNA in living cells for ratiometric fluorimetry of nucleic acid biomarker

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Abstract

A precisely designed dual-color biosensor has realized a visual assessment of thymidine kinase 1 (TK1) mRNA in both living cells and cell lysates. The oligonucleotide probe is constructed by hybridizing the antisense strand of the target and two recognition sequences, in which FAM serves as the donor and TAMRA as the acceptor. Once interacting with the target, two recognition strands are replaced, and then the antisense complementary sequence forms a more stable double-stranded structure. Due to the increasing spatial distance between two dyes, the FRET is attenuated, leading to a rapid recovery of FAM fluorescence and a reduction of TAMRA fluorescence. A discernible color response from orange to green could be observed by the naked eye, with a limit of detection (LOD) of 0.38 nM and 5.22 nM for spectrometer- and smartphone-based assays, respectively. The proposed ratiometric method transcends previous reports in its capacities in visualizing TK1 expression toward reliable nucleic acid biomarker analysis, which might establish a general strategy for ratiometric biosensing via strand displacement.

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All data generated or analyzed during this study are included in this published article and its supplementary information files.

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Funding

This work was supported by the National Natural Science Foundation of China (21703255), Anhui Province Science Fund for Outstanding Young Scholars (2208085Y27), Scientific Research Foundation for the Returned Overseas Chinese Scholars of Anhui Province-Key Projects (2021LCX003), Excellent Young Talents Fund Program of Higher Education Institutions of Anhui Province (gxyqZD2021102), Research Fund of Anhui Institute of Translational Medicine (2021zhyx-C17), Research Fund for the Doctoral Program of Anhui Medical University (XJ201808), and Research Fund for Scientific Research Level Improvement Plan of Anhui Medical University (2020xkjT001).

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Author notes

  1. Fengqi Dong and Weizhen Yan contributed equally to this work.

Authors and Affiliations

  1. School of Basic Medical Sciences, Biopharmaceutical Research Institute, Anhui Medical University, Hefei, Anhui, 230032, China

    Fengqi Dong, Mingsheng Qu, ** Zhang

  2. Department of Oncology, **angya Hospital, Central South University, Changsha, 410008, Hunan, China

    Weizhen Yan

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  1. Fengqi Dong
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Correspondence to Hanyuan Zhang, Wei Du or Tingting Zhao.

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This is an observational study. The Ethics Committee of Anhui Medical University has confirmed that no ethical approval is required.

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Dong, F., Yan, W., Qu, M. et al. Strand displacement–triggered FRET nanoprobe tracking TK1 mRNA in living cells for ratiometric fluorimetry of nucleic acid biomarker. Microchim Acta 191, 390 (2024). https://doi.org/10.1007/s00604-024-06453-7

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