Adaption of a Solid-State Nanopore to Homogeneous DNA Organization Verification and Label-Free Molecular Analysis without Covalent Modification
Adaption of a Solid-State Nanopore to Homogeneous DNA Organization Verification and Label-Free Molecular Analysis without Covalent Modification
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固态纳米孔无需共价修饰即可应用于同质 DNA 组织验证和无标记分子分析
DOI：
10.1021/acs.analchem.7b03442
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发表时间：
2018
影响因子：
7.4
通讯作者：
Li Bingling
中科院分区：
文献类型：
--
作者：
Zhu Zhentong;Zhou Ya;Xu Xiaolong;Wu Ruiping;Jin Yongdong;Li Bingling
最近的进展表明，通过控制寡核苷酸的热力学和动力学来增加核酸组织的设计。然而，这些DNA纳米技术的深入理解和进一步应用主要受到缺乏有效的分析方法的阻碍，这些分析方法足以调查它们。为了提供潜在的解决方案，我们进行了一项创新的探索，利用新兴的纳米孔技术在单分子水平和完全均匀的条件下，在没有共价修饰的情况下，表征DNA组织。借助计数和分析DNA组装结构通过锥形玻璃纳米孔（CGN）时易位诱导的电流下降，我们直接验证了由杂交链反应（HCR）模型产生的单个双螺旋串联体的形成。由于纳米孔技术的超灵敏度，那些在常规电泳图像上难以观察到的串联物被揭示出来。移位持续时间也提供了连接体的大致长度和折叠信息。这些优点被证明也适用于具有更复杂折叠行为的结构。最终，当与上游反应耦合时，CGN进一步成为一种通用检测器，甚至能够检测到其他核酸组织行为，以及无法产生巨大产物的靶标。这些结果有望推动纳米孔技术在核酸纳米技术领域的深入研究和应用。
Recent advances have shown increasing designs of nucleic acid organizations via controlling the thermodynamics and kinetics of oligonucleotides. Nevertheless, deeper understanding and further applications of these DNA nanotechnologies are majorly hampered by the lack of effective analytical methodologies that are competent enough to investigate them. To deliver a potential solution, here we developed an innovative exploration that employed the emerging nanopore technique to characterize DNA organization at the single-molecule level and in completely homogeneous condition without covalent modification. With the help of counting and profiling the translocation-induced current drop of a DNA assembly structure passing through a conical glass nanopore (CGN), we have directly verified the formation of the individual double-helix concatemer generated from our model, hybridization chain reaction (HCR). Due to the ultrasensitivity of the nanopore technology, those concatemers that were difficult to observe on a conventional electrophoresis image were brought to light. The translocation duration time also provided the approximate length and folding information for the concatemers. These advantages were proven also applicable to structures with more sophisticated folding behaviors. Eventually, when coupling with an upstream reaction, CGN was further turned to a universal detector that was capable of even detecting other nucleic acid organization behaviors as well as targets that were unable to generate huge products. All of these results are expected to promote deeper study and applications of the nanopore technique in the field of nucleic acid nanotechnology.