Fluorometric determination of RNase H via a DNAzyme conjugated to reduced graphene oxide, and its application to screening forinhibitors and activators

文献类型： 外文期刊

第一作者： Tong, Chunyi

作者： Tong, Chunyi;Zhou, Ting;Zhao, Chuan;Liu, Bin;Fan, Jialong;Li, Dan;Yuan, Liqun;Xu, Ying;Zhu, Aiguo

作者机构：

关键词： Fluorescent probe; Nanomaterial; Signal amplification; Quenching efficiency; Tumor cell lines; High sensitivity; Concentration-dependent manner; Model docking; Kinetic analysis; Hepatitis C virus

期刊名称：MICROCHIMICA ACTA （ 影响因子：5.833； 五年影响因子：5.357 ）

ISSN： 0026-3672

年卷期： 2019 年 186 卷 6 期

页码：

收录情况： SCI

摘要： A new fluorometric method is delineated for the detection of RNase H activity by combining DNAzyme with reduced graphene oxide (rGO). In the absence of RNase H, the fluorescence of FAM-labeled probe is quenched due to the strong adsorption on the rGO. The presence of RNase H can release the active DNAzyme from the DNA-RNA chimeric strand. This triggers the cleavage of the signal probe at the rA site with the help of the cofactor Mg2+. The recycle cleavage can directly result in the amplifiedsignal emitted bytheFAM-labeled short fragment. The method allows the activity of RNase H to be detected in a linear range of 0.01 to 5UmL(-1). The detection limit of 0.018UmL(-1) is calculated by the principle of three-time standard deviation over the blank signal. Then, RNase H-targeting natural compounds were screenedfor their inhibitory action. Among the investigated compounds, five were screened as RNase H inhibitors in a concentration-dependent manner, and 4 compounds were identified as activators. Finally, the method was reliably used for discriminating the difference of RNase H activity in human serum. It is found that RNase H activity was upregulated in patients with hepatitis C virus infection.

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