The effects of overhang placement and multivalency on cell labeling by DNA origami.
The effects of overhang placement and multivalency on cell labeling by DNA origami.
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DOI：
10.1039/d0nr09212f
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发表时间：
2021-04-14
期刊：
影响因子：
6.7
通讯作者：
Taylor RE
中科院分区：
文献类型：
--
作者：
Liu Y ;Wijesekara P ;Kumar S ;Wang W ;Ren X ;Taylor RE
通过与细胞膜的靶向结合，结构DNA纳米技术有可能引导和固定生物分子，如药物，生长因子和纳米生物传感器到细胞表面。在这项研究中，我们研究了三种不同的DNA折纸形状通过胆固醇锚定培养的内皮细胞的靶向结合效率。我们的研究结果表明，标记效率是高度依赖于折纸的形状以及结合突出端的数量和位置。通过均匀的表面结合悬突间距，三维各向同性纳米球和一维各向异性纳米棒有效地标记细胞，并且各向同性纳米球标记与独立的结合模型拟合良好。进行各向异性纳米瓦片的面装饰和边缘装饰以研究结合悬突位置对细胞标记的影响，并且仅边缘装饰的纳米瓦片成功地标记细胞。边缘邻近性研究表明，可以通过分别向边缘和顶点移动结合突出端来调制纳米片和纳米棒两者中的标记效率。此外，我们证明，虽然双链DNA（dsDNA）桥系链可以挽救的表面装饰的矩形板的标记效率，这种效果也取决于桥系链的边缘或顶点的纳米结构的接近。所有三种纳米形状的最终比较显示，末端标记的纳米棒和纳米球实现了最高的绝对标记强度，但最高的信噪比，计算为整体标记与无引发剂背景标记的比率，是由末端标记的纳米棒实现的，边缘标记的纳米片排在第二位，略高于纳米球。这项研究的发现可以帮助我们进一步了解影响胆固醇锚定膜附着的因素，从而为未来功能性DNA纳米结构的合理设计提供指导。
Through targeted binding to the cell membrane, structural DNA nanotechnology has the potential to guide and affix biomolecules such as drugs, growth factors and nanobiosensors to the surfaces of cells. In this study, we investigated the targeted binding efficiency of three distinct DNA origami shapes to cultured endothelial cells via cholesterol anchors. Our results showed that the labeling efficiency is highly dependent on the shape of the origami as well as the number and the location of the binding overhangs. With a uniform surface spacing of binding overhangs, 3D isotropic nanospheres and 1D anisotropic nanorods labeled cells effectively, and the isotropic nanosphere labeling fit well with an independent binding model. Face-decoration and edge-decoration of the anisotropic nanotile were performed to investigate the effects of binding overhang location on cell labeling, and only the edge-decorated nanotiles were successful at labeling cells. Edge proximity studies demonstrated that the labeling efficiency can be modulated in both nanotiles and nanorods by moving the binding overhangs towards the edges and vertices, respectively. Furthermore, we demonstrated that while double-stranded DNA (dsDNA) bridge tethers can rescue the labeling efficiency of the face-decorated rectangular plate, this effect is also dependent on the proximity of bridge tethers to the edges or vertices of the nanostructures. A final comparison of all three nanoshapes revealed that the end-labeled nanorod and the nanosphere achieved the highest absolute labeling intensities, but the highest signal-to-noise ratio, calculated as the ratio of overall labeling to initiator-free background labeling, was achieved by the end-labeled nanorod, with the edge-labeled nanotile coming in second place slightly ahead of the nanosphere. The findings from this study can help us further understand the factors that affect membrane attachment using cholesterol anchors, thus providing guidelines for the rational design of future functional DNA nanostructures.
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