Quantification of Strand Accessibility in Biostable DNA Origami with Single-Staple Resolution,ACS Nano

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Quantification of Strand Accessibility in Biostable DNA Origami with Single-Staple Resolution
ACS Nano ( IF 15.8 ) Pub Date : 2021-10-06 , DOI: 10.1021/acsnano.1c05540
Alexandra S Eklund _{1,

2} , Alice Comberlato ₃ , Ian A Parish _{4,

5} , Ralf Jungmann _{1,

2} , Maartje M C Bastings _{3,

6}

Affiliation

DNA-based nanostructures are actively gaining interest as tools for biomedical and therapeutic applications following the recent development of protective coating strategies prolonging structural integrity in physiological conditions. For tailored biological action, these nanostructures are often functionalized with targeting or imaging labels using DNA base pairing. Only if these labels are accessible on the structure’s surface will they be able to interact with their intended biological target. However, the accessibility of functional sites for different geometries and environments, specifically after the application of a protective coating, is currently not known. Here, we assay this accessibility on the level of single handle strands with two- and three-dimensional resolution using DNA-PAINT and show that the hybridization kinetics of top and bottom sides on the same nanostructure linked to a surface remain unaltered. We furthermore demonstrate that the functionality of the structures remains available after an oligolysine-PEG coating is applied, enabling bioassays where functionality and stability are imperative.

中文翻译：

用单钉分辨率量化生物稳定 DNA 折纸中的链可及性

随着最近开发的保护涂层策略延长生理条件下的结构完整性，基于 DNA 的纳米结构作为生物医学和治疗应用的工具正在积极获得关注。对于定制的生物作用，这些纳米结构通常使用 DNA 碱基配对通过靶向或成像标记进行功能化。只有当这些标签在结构表面可触及时，它们才能与预期的生物靶标相互作用。然而，不同几何形状和环境的功能位点的可访问性，特别是在应用保护涂层之后，目前尚不清楚。这里，我们使用 DNA-PAINT 在具有二维和三维分辨率的单柄链水平上分析了这种可及性，并表明连接到表面的同一纳米结构上的顶部和底部的杂交动力学保持不变。我们进一步证明，在应用寡聚赖氨酸-PEG 涂层后，结构的功能仍然可用，从而能够在功能和稳定性必不可少的情况下进行生物测定。

更新日期：2021-11-23

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