A Novel DNAzyme Signal Amplification-based Colorimetric Method for RNase H Assays

• Published in: Analytical Sciences Vol. 37; no. 12; pp. 1675 - 1680
• Main Authors: XIE, Ye, ZHANG, Sina, DENG, Ting, ZHANG, Ke, REN, Jiali, LI, Jishan
• Format: Journal Article
• Language: English
• Published: Singapore The Japan Society for Analytical Chemistry 10-12-2021; Springer Nature Singapore; Japan Science and Technology Agency
• Subjects: Amplification; Analytical Chemistry; Chemistry; colorimetric; Colorimetry; Deoxyribonucleic acid; DNA; DNA, Catalytic - metabolism; DNAzyme; G-quadruplex/hemin; G-Quadruplexes; Hemin; Horseradish peroxidase; Lysates; Mimicry; Nucleotide sequence; Original Papers; Oxidation; Peroxidase; Ribonuclease H; Ribonucleic acid; RNA; RNase H; Selectivity; signal amplification; Substrates; Sulfonic acid; Visual signals; DNAzyme; G-quadruplex/hemin; colorimetric; RNase H; signal amplification
• ISSN: 0910-6340; 1348-2246
• DOI: 10.2116/analsci.20P337

A simple visual strategy was developed for the RNase H colorimetric measurement using DNAzyme-mediated signal amplification. When RNase H was presented, the RNA strand of the duplex formed by the G-rich DNA sequence (G-Rich) and its complementary RNA sequence (cp-RNA) was digested, releasing G-Rich to form HRP-mimicking DNAzymes of the G-quadruplex/hemin complexes in the presence of hemin. These DNAzymes catalyze the oxidation reaction of the substrate of 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) to produce green-color products of ABTS•−, allowing for the detection of RNase H. A horseradish peroxidase (HRP)-mimicking DNAzyme of the G-quadruplex/hemin complex was used to mediate the signal amplification in the sensing strategy, resulting in high selectivity and sensitivity. This proposed colorimetric method shows a low detection limit of 0.04 U/mL, with a detection range of 0.1 to 3 U/mL. Moreover, this colorimetric method has been successfully used for RNase H assays in complicated biosamples, such as cell lysates. These results indicate that our colorimetric method not only detects RNase H in an ideal system, but also in real samples.