Department of Cellular and Molecular Biophysics, Max Planck Institute of Biochemistry, Am Klopferspitz 18, D-82152 Martinsried, Germany.
Exzellenzcluster ORIGINS, Boltzmannstrasse 2, D-85748 Garching, Germany.
J Phys Chem B. 2021 Dec 9;125(48):13181-13191. doi: 10.1021/acs.jpcb.1c07694. Epub 2021 Nov 24.
Nanotechnology often exploits DNA origami nanostructures assembled into even larger superstructures up to micrometer sizes with nanometer shape precision. However, large-scale assembly of such structures is very time-consuming. Here, we investigated the efficiency of superstructure assembly on surfaces using indirect cross-linking through low-complexity connector strands binding staple strand extensions, instead of connector strands binding to scaffold loops. Using single-molecule imaging techniques, including fluorescence microscopy and atomic force microscopy, we show that low sequence complexity connector strands allow formation of DNA origami superstructures on lipid membranes, with an order-of-magnitude enhancement in the assembly speed of superstructures. A number of effects, including suppression of DNA hairpin formation, high local effective binding site concentration, and multivalency are proposed to contribute to the acceleration. Thus, the use of low-complexity sequences for DNA origami higher-order assembly offers a very simple but efficient way of improving throughput in DNA origami design.
纳米技术通常利用 DNA 折纸纳米结构组装成更大的超结构，尺寸可达微米级，形状精度达到纳米级。然而，这种结构的大规模组装非常耗时。在这里，我们研究了通过低复杂度连接器链通过与订书钉链延伸结合而非与支架环结合进行间接交联在表面上进行超结构组装的效率。使用包括荧光显微镜和原子力显微镜在内的单分子成像技术，我们表明低序列复杂度连接器链允许 DNA 折纸超结构在脂质膜上形成，超结构的组装速度提高了一个数量级。提出了许多效应，包括抑制 DNA 发夹形成、高局部有效结合位点浓度和多价性，以促进加速。因此，使用低复杂度序列进行 DNA 折纸更高阶组装为提高 DNA 折纸设计的通量提供了一种非常简单但有效的方法。
