Self-assembled DNA origami-based duplexed aptasensors combined with centrifugal filters for efficient and rechargeable ATP detection.
Self-assembled DNA origami-based duplexed aptasensors combined with centrifugal filters for efficient and rechargeable ATP detection.
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DOI：
10.1016/j.bios.2022.114336
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发表时间：
2022-05
影响因子：
12.6
通讯作者：
Xiaojuan Wang;Shuning Li;Ruipeng Chen;Zefeng Mao;Shuyue Ren;Jun Liang;Zhixian Gao
中科院分区：
文献类型：
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作者：
Xiaojuan Wang;Shuning Li;Ruipeng Chen;Zefeng Mao;Shuyue Ren;Jun Liang;Zhixian Gao
DNA折纸技术在生物传感器应用方面具有巨大潜力。在这里，我们描述了用于荧光适体精确定位的自组装 DNA 折纸生物传感器的构建。由于DNA折纸和适体之间的分子量差异，采用离心过滤器来定量检测三磷酸腺苷（ATP）。选择荧光报道分子6-羧基荧光素FAM标记的ATP特异性适配体（FAM-适配体）作为识别元件和信号探针。 ATP 双链适体通过碱基互补配对与三角形 DNA 折纸结合，从而在折纸阵列上产生高荧光信号。 ATP与FAM-适体的竞争性结合触发了FAM-适体-ATP复合物从折纸阵列表面的释放，导致荧光信号减弱。对于 ATP 定量，采用 100 kD 离心过滤器，然后测量过滤器装置的折纸阵列上捕获的荧光信号。通过原子力显微镜、激光共焦显微镜和电泳对折纸适体阵列的成功合成进行了表征。荧光测量与 ATP 浓度的对数在 0.1–100 ng mL−1 范围内表现出良好的线性关系，检测限为 0.29 ng mL−1。通过替换适体和互补链，我们证明了该方法用于 17β-雌二醇检测的潜力。考虑到检测机制是基于DNA链的杂交和置换，该检测系统具有充电的潜力。我们的研究为 DNA 折纸技术在小分子检测中的应用提供了新的见解。
DNA origami technology has great potential for biosensor applications. Here, we described the construction of a self-assembled DNA origami biosensor for the precise localization of fluorescent aptamers. Due to the molecular weight difference between DNA origami and aptamer, centrifugal filters were used to quantitatively detect adenosine triphosphate (ATP). The ATP-specific aptamer labeled with fluorescence reporter 6-carboxyfluorescein FAM (FAM-aptamer) was selected as the recognition element and signal probe. ATP duplexed aptamers bound to triangular DNA origami by base-complementary pairing, resulting in high fluorescence signals on the origami arrays. The competitive binding of ATP toward the FAM-aptamer triggered the release of FAM-aptamer-ATP complexes from the surface of the origami array, resulting in weakened fluorescence signals. For ATP quantification, 100 kD centrifugal filters were employed, followed by measurement of the fluorescence signal trapped on the origami arrays of the filter device. The successful synthesis of origami-aptamer arrays was characterized by atomic force microscopy, laser confocal microscopy, and electrophoresis. Fluorescence measurements exhibited an excellent linear relationship with logarithms of ATP concentrations within 0.1–100 ng mL−1, with a detection limit of 0.29 ng mL−1. By replacing aptamers and complementary strands, we demonstrated the potential of this method for 17β-estradiol detection. Considering that the detection mechanism is based on the hybridization and displacement of DNA strands, the detection system had the potential for recharging. Our study provides new insights into applying DNA origami technology in small molecule detection.