Probing Cellular Molecules with PolyA-Based Engineered Aptamer Nanobeacon
Probing Cellular Molecules with PolyA-Based Engineered Aptamer Nanobeacon
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DOI：
10.1021/acsami.6b16764
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发表时间：
2017-03-08
影响因子：
9.5
通讯作者：
Pei, Hao
中科院分区：
文献类型：
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作者：
Chen, Lizhen;Chao, Jie;Pei, Hao
三磷酸腺苷（ATP）是一种在许多细胞过程中至关重要的中心代谢产物。发展灵敏和选择性的体内ATP水平检测方法在诊断和治疗诊断应用中至关重要。在这项工作中，我们开发了一种基于polyA的适配体纳米信标（PAaptNB），提高了ATP分析的效率和速度。我们发现，PAaptNB的解离常数和竞争性结合动力学可以通过调节polyA的长度来可编程地调节。当polyA长度达到30个碱基时，可以实现用PAaptNB进行ATP测定的10 μ M检测限（与常规的基于硫醇的适体纳米信标相比，类似于10倍的改进）。通过对细胞内ATP分子进行成像，进一步证明了PAaptNB用于体内测定的可行性。本研究为构建高效、快速的细胞分子生物传感器提供了一种新的思路，在生物分析和诊断治疗中具有广阔的应用前景。
Adenosine triphosphate (ATP) is a central metabolite that is of critical importance in many cellular processes. The development of sensitive and selective methods for the detection of ATP level in vivo is crucial in diagnostic and theranostic applications. In this work, we have developed a polyA-based aptamer nanobeacon (PAaptNB) with improved efficiency and speed of ATP analysis. We found that the dissociation constants and competitive binding kinetics of the PAaptNB could be programmably regulated by adjusting the polyA length. When the polyA length reached to 30 bases, a 10 mu M detection limit for ATP assay with PAaptNB can be achieved (similar to 10-fold improvement compared with the conventional thiol-based aptamer nanobeacon). The feasibility of the PAaptNB for in vivo assay was further demonstrated by imaging intracellular ATP molecules. This study provides a new strategy to construct high-efficiency and high-speed biosensors for cellular molecules analysis, which holds great potential in bioanalysis and theranostic applications.