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


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

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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).

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  • Multiple G-triplex
  • ABTS
  • H2O2
  • DNAzyme
  • Isothermal amplification