Abstract
DNA G-quadruplexes are special three-dimensional (3D) DNA nanostructures formed by specific G-rich DNA sequences. These 3D DNA nanostructures can bind with hemin and significantly improve the intrinsic peroxidase activity of hemin. Besides this function, they also enhance the fluorescence intensity of some G-quadruplex-specific dyes. Owing to these features, G-quadruplexes possess several superiorities in the detection of enzymes involved in nucleic acid metabolism, including facile probe fabrication without labeling, simple detection process without washing or separation steps, rapid observation by naked eyes, and easy integration with nucleic acid amplification strategies to amplify signals. Herein, we describe two strategies for label-free detection of enzyme activity based on DNA G-quadruplexes. To increase sensitivity, template-dependent and template-independent DNA amplifications were introduced for the amplification of G-rich DNA sequences. DNA methyltransferase and terminal deoxynucleotidyl transferase were detected as two model analytes, respectively.
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Acknowledgments
This work was funded by the National Natural Science Foundation of China (Nos. 21222507, 21175036, 21235002, 21575038, and 21305037), the Foundation for Innovative Research Groups of NSFC (Grant 21221003), and the Natural Science Foundation of Hunan Province (No. 2015JJ1005).
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Liu, Z. et al. (2017). DNA G-Quadruplex-Based Assay of Enzyme Activity. In: Ke, Y., Wang, P. (eds) 3D DNA Nanostructure. Methods in Molecular Biology, vol 1500. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6454-3_10
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DOI: https://doi.org/10.1007/978-1-4939-6454-3_10
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