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A colorimetric zinc(II) assay based on the use of hairpin DNAzyme recycling and a hemin/G-quadruplex lighted DNA nanoladder

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Abstract

A method is described for sensitive colorimetric determination of zinc(II) ions that uses (a) a Zn(II)-responsive hairpin DNAzyme that assists target recycling, (b) hybridization chain reaction, and (c) hemin/G-quadruplex DNA nanoladder. The Zn(II)-responsive split of the hairpin DNAzyme (HD) acts as the recognition and transformation probe. Upon addition of Zn (II) and enzyme strand, a duplex is formed in the loop region of hairpin. The caged initiator sequence is subsequently liberated from the HD by the Zn(II)-selective split of the substrate strand. This cleavage induces an enzyme strand recycling for the next round of cleavage. As a result, the initiator DNA is accumulated and cross-opened H1 and H2 to start a hybridization chain reaction (HCR). The caged G-quadruplex is released after the HCR to recruit hemin to form the hemin/G-quadruplex that is inserted into the DNA nanoladder. Once formed in the DNA nanoladder, these act as catalytic labels for the ABTS, resulting a green color change. This cascade amplification strategy allows 10 nM to 100 μM of Zn(II) to be linearly quantified by colorimetry at 415 nm with a detection limit of 3.5 nM. The recoveries ranged from 97.7 to 108.3% were obtained, confirming high reliability of the method for Zn2+ analysis in lake water samples.

A colorimetric assay for Zn2+ using hairpin DNAzyme (HD) assistant cycle and hemin/G-quadruplex lighted nanoladders is designed. A Zn2+-responsive split of HD is designed as the recognition and transformation probe. The hemin/G-Quadruplex inserted nanoladder act as reporter for signal readout.

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Acknowledgments

This work is financially supported by the major science and technology project to create new crop varieties using gene transfer technology” (2018ZX0801102B-002) and the National Natural Science Foundation of China (No.31801626, No.31871875).

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Authors and Affiliations

  1. Bei**g Advanced Innovation Center for Food Nutrition and Human Health, College of Food Science & Nutritional Engineering, China Agricultural University, Bei**g, 100083, China

    Longjiao Zhu, **angli Shao, Kunlun Huang, Yunbo Luo & Wentao Xu

  2. Key Laboratory of Safety Assessment of Genetically Modified Organism (Food Safety), Ministry of Agriculture, Bei**g, 100083, China

    Longjiao Zhu, Guishan Li, **angli Shao, Kunlun Huang, Yunbo Luo & Wentao Xu

  3. Bei**g Laboratory for Food Quality and Safety, College of Food Science and Nutritional Engineering, China Agricultural University, Bei**g, 100083, China

    Kunlun Huang & Wentao Xu

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  1. Longjiao Zhu
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  2. Guishan Li
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  4. Kunlun Huang
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Correspondence to Wentao Xu.

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Zhu, L., Li, G., Shao, X. et al. A colorimetric zinc(II) assay based on the use of hairpin DNAzyme recycling and a hemin/G-quadruplex lighted DNA nanoladder. Microchim Acta 187, 26 (2020). https://doi.org/10.1007/s00604-019-3996-2

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