DNA 纳米结构的相关超分辨率和原子力显微镜以及可寻址位点缺陷的表征,ACS Nano

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DNA 纳米结构的相关超分辨率和原子力显微镜以及可寻址位点缺陷的表征
ACS Nano ( IF 15.8 ) Pub Date : 2021-06-17 , DOI: 10.1021/acsnano.1c01976
Christopher M Green _{1,

2} , William L Hughes , Elton Graugnard , Wan Kuang

Affiliation

为了使 DNA 纳米结构在现实世界中的应用取得成果，需要先进的显微镜技术来阐明限制可寻址位点可用性的因素。关联显微术将两个或多个显微术结合起来表征同一样品，是一种克服单个技术局限性的方法，但它在 DNA 纳米技术中的应用有限。我们开发了一种可访问的策略，用于高分辨率、基于相关 DNA 的点积累，用于 DNA 纳米结构的纳米级地形图 (DNA-PAINT) 超分辨率和原子力显微镜 (AFM) 成像，通过一种简单而稳健的方法选择性结合DNA 折纸盖玻璃。使用这种技术，我们检查了 DNA 折纸上可寻址的“对接”位点，以区分两种缺陷情况——结构上并入但不活动的对接点，以及未合并的对接点。我们发现超过 75% 的有缺陷的停靠站点已合并但处于非活动状态，这表明未合并的链在限制可寻址站点的可用性方面发挥了次要作用。我们进一步探讨了链纯化、紫外线照射和光氧化对可用性的影响，提供了关于潜在缺陷来源和提高 DNA 纳米结构保真度途径的见解。

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Correlative Super-Resolution and Atomic Force Microscopy of DNA Nanostructures and Characterization of Addressable Site Defects

To bring real-world applications of DNA nanostructures to fruition, advanced microscopy techniques are needed to shed light on factors limiting the availability of addressable sites. Correlative microscopy, where two or more microscopies are combined to characterize the same sample, is an approach to overcome the limitations of individual techniques, yet it has seen limited use for DNA nanotechnology. We have developed an accessible strategy for high resolution, correlative DNA-based points accumulation for imaging in nanoscale topography (DNA-PAINT) super-resolution and atomic force microscopy (AFM) of DNA nanostructures, enabled by a simple and robust method to selectively bind DNA origami to cover glass. Using this technique, we examined addressable “docking” sites on DNA origami to distinguish between two defect scenarios–structurally incorporated but inactive docking sites, and unincorporated docking sites. We found that over 75% of defective docking sites were incorporated but inactive, suggesting unincorporated strands played a minor role in limiting the availability of addressable sites. We further explored the effects of strand purification, UV irradiation, and photooxidation on availability, providing insight on potential sources of defects and pathways toward improving the fidelity of DNA nanostructures.

更新日期：2021-07-27

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