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CRISPR-Cas13a-Triggered DNAzyme Signal Amplification-Based Colorimetric miRNA Detection Method and Its Application in Evaluating the Anxiety

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Abstract

The development of a bio-sensing strategy based on CRISPR/Cas that is exceptionally sensitive is crucial for the identification of trace molecules. Colorimetric miRNA detection utilizing CRISPR/Cas13a-triggered DNAzyme signal amplification was described in this article. The developed strategy was implemented for miRNA-21 detection as a proof of concept. The cleavage activity of Cas13a was triggered when the target molecule bonded to the Cas13a-crRNA complex and cleaved uracil ribonucleotides (rU) in the substrate probe. As a consequence, the S chain was liberated from the T chain that had been modified on magnetic beads (MB). The G-rich sections were then exposed when the catalytic hairpin assembly between the H1 and H2 probes was activated by the released T@MB. G-rich section can fold into G-quadruplex. By catalyzing the formation of green ABTS3– via HRP-mimicking G-quadruplex/hemin complexes, colorimetric measurements of miRNA can be achieved visually through DNAzyme-mediated signal amplification. The method demonstrated a low limit of detection of 27 fM and a high selectivity towards target miRNA eventually. As a result, the developed strategy provides a clinical application platform for the detection of miRNAs that is both ultrasensitive and extremely specific.

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Data Availability

All data generated and analyzed during this study are included in this article.

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Funding

The research was financially supported by the Hengshui City Science and Technology Plan Project (project number: 2023014010Z).

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Authors and Affiliations

  1. Department of Stomatology, Hengshui People’s Hospital Affiliated to Hebei Medical University, No. 180 Renmin East Road, Hengshui City, 053000, Hebei Province, China

    Na Yan, Zhiwei Hu & Lingling Zhang

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  1. Na Yan
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  2. Zhiwei Hu
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  3. Lingling Zhang
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Contributions

Z.Y. is the supervisor of the team in all research steps including designing, data analysis, and manuscript writing. N.Y. has the main role for experimental data collection, data gathering, preparation of results, and data analysis. Z.H. and L.Z. assisted in the data analysis.

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Correspondence to Na Yan.

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Yan, N., Hu, Z. & Zhang, L. CRISPR-Cas13a-Triggered DNAzyme Signal Amplification-Based Colorimetric miRNA Detection Method and Its Application in Evaluating the Anxiety. Appl Biochem Biotechnol (2024). https://doi.org/10.1007/s12010-024-04951-1

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