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基于 G-四链体自组装的超灵敏鸟嘌呤线共振光散射法测定 microRNA-122。

An ultrasensitive guanine wire-based resonance light scattering method using G-quadruplex self-assembly for determination of microRNA-122.

机构信息

Key Laboratory for Green Organic Synthesis and Application of Hunan Province, Key Laboratory of Environmentally Friendly Chemistry and Application of Ministry of Education, College of Chemistry, Xiangtan University, Xiangtan, 411105, China.

College of Science, Hunan Agricultural University, Changsha, 410128, China.

出版信息

Mikrochim Acta. 2019 Aug 3;186(9):599. doi: 10.1007/s00604-019-3707-z.

Abstract

An enzyme-free resonance light scattering (RLS) method is described for the determination of microRNA-122. A guanine nanowire (G-wire) is used that consists of a predesigned DNA1 and a G-quadruplex sequence DNA2. These hybridize with microRNA-122 and partially hybridize with DNA2. After formation of stable double strands with DNA1, DNA2 is released. On addition of K and Mg ions, the G-quadruplex sequences undergo self-assembly to form long filamentous G-wires. This increases the intensity of RLS. A 6.1 pM detection limit was obtained, and the linear response covers the 50 pM to 300 nM microRNA concentration range. The method was successfully applied to the quantitation of microRNA-122 in hepatocellular carcinoma cell lysates. Conceivably, this assay can be extended to other RLS methods for biomarker detection by simply changing the sequence of DNA1. Graphical abstract The G-quadruplex sequences of DNA2 were locked with DNA1. The G-quadruplex fragments of DNA2 were released after the hybridization of microRNA-122 with DNA1. These liberated G-quadruplex sequences were self-assembled into long filamentous guanine nanowires (G-wires) which increased resonance light intensity in the presence of Mg.

摘要

一种无酶共振光散射(RLS)方法被用来测定 microRNA-122。该方法使用了一条由预先设计的 DNA1 和 G-四链体序列 DNA2 组成的鸟嘌呤纳米线(G-wire)。这些 DNA 与 microRNA-122 杂交,并与 DNA2 部分杂交。与 DNA1 形成稳定的双链后,释放出 DNA2。加入 K 和 Mg 离子后,G-四链体序列自行组装形成长丝状 G-wire。这增加了 RLS 的强度。该方法的检测限为 6.1 pM,线性响应范围涵盖 50 pM 至 300 nM 的 microRNA 浓度。该方法成功应用于肝癌细胞裂解物中 microRNA-122 的定量。可以想象,通过简单改变 DNA1 的序列,这种测定方法可以扩展到其他用于生物标志物检测的 RLS 方法。

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