State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Key Laboratory for Bio-Nanotechnology and Molecular Engineering of Hunan Province, Hunan University , Changsha 410082, People's Republic of China.
Anal Chem. 2017 Mar 21;89(6):3590-3596. doi: 10.1021/acs.analchem.6b04918. Epub 2017 Mar 10.
DNA nanostructures have emerged as powerful and versatile building blocks for the construction of programmable nanoscale structures and functional sensors for biomarker detection, disease diagnostics, and therapy. Here we integrated multiple sensing modules into a single DNA three-dimensional (3D) nanoarchitecture with a triangular-prism (TP) structure for ratiometric and multiplexed biomolecule detection on a single microbead. In our design, the complementary hybridization of three clip sequences formed TP nanoassemblies in which the six single-strand regions in the top and bottom faces act as binding sites for different sensing modules, including an anchor module, reference sequence module, and capture sequence module. The multifunctional modular TP nanostructures were thus exploited for ratiometric and multiplexed biomolecule detection on microbeads. Microbead imaging demonstrated that, after ratiometric self-calibration analysis, the imaging deviations resulting from uneven fluorescence intensity distribution and differing probe concentrations were greatly reduced. The rigid nanostructure also conferred the TP as a framework for geometric positioning of different capture sequences. The inclusion of multiple targets led to the formation of sandwich hybridization structures that gave a readily detectable optical response at different fluorescence channels and distinct fingerprint-like pattern arrays. This approach allowed us to discriminate multiplexed biomolecule targets in a simple and efficient fashion. In this module-designed strategy, the diversity of the controlled DNA assembly coupled with the geometrically well-defined rigid nanostructures of the TP assembly provides a flexible and reliable biosensing approach that shows great promise for biomedical applications.
DNA 纳米结构已成为构建可编程纳米结构和功能传感器的强大而多功能的构建模块，可用于生物标志物检测、疾病诊断和治疗。在这里，我们将多个传感模块集成到具有三角形棱镜 (TP) 结构的单个 DNA 三维 (3D) 纳米结构中，用于在单个微珠上进行比率和多重生物分子检测。在我们的设计中，三个夹序列的互补杂交形成了 TP 纳米组装体，其中顶部和底部表面的六个单链区域充当不同传感模块的结合位点，包括锚定模块、参考序列模块和捕获序列模块。因此，多功能模块化 TP 纳米结构可用于在微珠上进行比率和多重生物分子检测。微珠成像表明，在进行比率自校准分析后，由于荧光强度分布不均匀和探针浓度不同而导致的成像偏差大大降低。刚性纳米结构还赋予 TP 作为不同捕获序列的几何定位框架。包含多个靶标会导致形成夹心杂交结构，在不同荧光通道和独特的指纹样图案阵列中产生可检测的光学响应。这种方法允许我们以简单有效的方式区分多重生物分子靶标。在这种模块设计策略中，受控制的 DNA 组装的多样性与 TP 组装的几何形状明确的刚性纳米结构相结合，提供了一种灵活可靠的生物传感方法，在生物医学应用中具有广阔的应用前景。
