A self-assembling RNA aptamer-based graphene oxide sensor for the turn-on detection of theophylline in serum
A self-assembling RNA aptamer-based graphene oxide sensor for the turn-on detection of theophylline in serum
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基于自组装RNA适体的氧化石墨烯传感器，用于启动检测血清中的茶碱
DOI：
10.1016/j.bios.2016.06.024
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发表时间：
2016-12-15
影响因子：
12.6
通讯作者：
Li, Fu-Rong
中科院分区：
文献类型：
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作者：
Ling, Kai;Jiang, Hongyan;Li, Fu-Rong
迄今为止，已经开发出几种基于RNA适体的有效荧光生物传感器，因为RNA在核酸酶存在下的固有不稳定性排除了RNA适体用于生物流体分析的应用。在这项研究中，我们开发了一个简单的，灵敏的，选择性的打开荧光适体检测血清中的茶碱，利用配体诱导的自组装RNA适体和两个不同的相互作用阶段的适体片段与氧化石墨烯（GO）。茶碱RNA适体的单链（33聚体）在末端环区域处分裂成两个较短的片段，其中一个片段用荧光团（FAM）标记。在不存在茶碱的情况下，两个单独片段在GO上的吸附使荧光团非常接近GO表面，导致荧光的高效淬灭。该系统显示出非常低的背景荧光。相反，片段自组装成RNA适体/茶碱复合物，并从GO解离。猝灭的荧光显着恢复，茶碱可以在很宽的浓度范围从1到100 μ M，检测限为0.155 μ M和良好的选择性血清中检测。此外，由于RNA片段较短以及GO有效保护核酸酶切割的能力，RNA序列在实验过程中保持稳定。这种设计可以作为RNA适体传感器在临床环境中应用的一个例子。(C)2016爱思唯尔B. V.保留所有权利。
To date, few effective fluorescent biosensors based on RNA aptamers have been developed because the intrinsic instability of RNA in the presence of nucleases precludes the application of RNA aptamers for the analysis of biological fluids. In this study, we developed a simple, sensitive, selective turn-on fluorescent aptasensor for theophylline detection in serum, utilizing ligand-induced self-assembling RNA aptamers and two different interaction stages of the aptamer fragments with graphene oxide (GO). A single strand of the theophylline RNA aptamer (33-mer) was split at the end loop region into two shorter fragments, one of which was labeled with a fluorophore (FAM). In the absence of theophylline, the adsorption of the two individual fragments on GO brought the fluorophore in close proximity to the GO surface, resulting in highly efficient quenching of fluorescence. The system showed very low background fluorescence. Conversely, the fragments self-assembled into an RNA aptamer/theophylline complex and were dissociated from GO. The quenched fluorescence was significantly recovered, and theophylline could be detected at a wide range of concentrations from 1 to 100 mu M, with a detection limit of 0.155 mu M and good selectivity in serum. Moreover, because of the shorter RNA fragments and the effective protection ability of GO from nuclease cleavage, the RNA sequences remained stable during the experiments. This design may serve as an example for the application of RNA aptasensors in the clinical setting. (C) 2016 Elsevier B.V. All rights reserved.