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Sensitive colorimetric determination of microRNA let-7a through rolling circle amplification and a peroxidase-mimicking system composed of trimeric G-triplex and hemin DNAzyme

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Abstract

The authors have incidentally found that the three tandem repeats of a 13-mer G-rich oligomer (with sequence 5′-TGG GAA GGG AGG G-3′; referred to as G3) can directly fold into a stable G3 trimer. The G3 trimer/hemin DNAzyme exhibits an about 3-fold higher peroxidase-mimicking activity compared to the conventional G3/hemin DNAzyme. Combining this finding with rolling circle amplification (RCA), a colorimetric assay was developed for sensitive and specific determination of microRNA. In this method, each cycle of RCA generates three catalytic units. This leads to a significant signal amplification of the RCA. Using let-7a as a model analyte, the colorimetric method (best performed at 420 nm) exhibits high sensitivity toward microRNA-let-7a with a 37 fM detection limit and an analytical range that covers 3 orders of magnitude. The method was applied to the determination of let-7a in some cell lysates.

This G-triplex trimer-based rolling circle amplification (RCA) method can produce three catalytic units per RCA cycle, which can significantly improve the amplification efficiency of RCA.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (No. 21775099 and 21974082).

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Authors and Affiliations

  1. Key Laboratory of Analytical Chemistry for Life Science of Shaanxi Province School of Chemistry & Chemical Engineering, Shaanxi Normal University, Xi’an, 710062, China

    Ruiying Li, Qiang Liu, Yan Jin & Baoxin Li

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  1. Ruiying Li
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  2. Qiang Liu
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  3. Yan Jin
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  4. Baoxin Li
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Correspondence to Baoxin Li.

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Li, R., Liu, Q., Jin, Y. et al. Sensitive colorimetric determination of microRNA let-7a through rolling circle amplification and a peroxidase-mimicking system composed of trimeric G-triplex and hemin DNAzyme. Microchim Acta 187, 139 (2020). https://doi.org/10.1007/s00604-019-4093-2

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  • DOI: https://doi.org/10.1007/s00604-019-4093-2

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