Gigadalton-scale shape-programmable DNA assemblies
Gigadalton-scale shape-programmable DNA assemblies
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DOI：
10.1038/nature24651
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发表时间：
2017-12-07
期刊：
影响因子：
64.8
通讯作者：
Dietz, Hendrik
中科院分区：
文献类型：
--
作者：
Wagenbauer, Klaus F.;Sigl, Christian;Dietz, Hendrik
天然生物分子组装体，如分子马达、酶、病毒和亚细胞结构，通常通过多个亚基的自限性分级寡聚化形成（1-3）。通过结合分级组装和对称性，大型结构也可以从几个组件有效组装，这是病毒衣壳的一种策略（4）。从头蛋白质设计（5-9）和RNA（10，11）和DNA纳米技术（12-14）旨在模拟这些能力，但自下而上构建具有病毒和其他亚细胞组分尺寸和复杂性的人工结构仍然具有挑战性。在这里，我们展示了自然组装原理可以与DNA折纸（15-24）的方法相结合，以产生具有受控尺寸的千兆道尔顿级结构。DNA序列信息用于编码单个DNA折纸构建块的形状，这些构建块之间相互作用的几何形状和细节然后控制它们在高阶组装中的拷贝数、位置和方向。我们通过创建直径高达350纳米的平面环和原子质量高达330兆道尔顿，微米长，厚管的大小与一些杆菌相称，以及尺寸高达1.2千兆道尔顿和直径450纳米的三维多面体组件来说明这一策略。我们实现了高效的组装，产量高达90%，通过使用具有验证结构和足够刚度的积木，以及具有相互作用图案的精确设计，确保分层组装是自限性的，并且能够在平衡状态下进行，以允许纠错。我们希望我们的方法，它使自组装的结构与病毒和细胞器的大小接近，可以很容易地用于创建一系列其他复杂的结构与明确定义的大小，通过利用模块化和高度的可寻址性的DNA折纸积木。
Natural biomolecular assemblies such as molecular motors, enzymes, viruses and subcellular structures often form by self-limiting hierarchical oligomerization of multiple subunits(1-3). Large structures can also assemble efficiently from a few components by combining hierarchical assembly and symmetry, a strategy exemplified by viral capsids(4). De novo protein design(5-9) and RNA(10,11) and DNA nanotechnology(12-14) aim to mimic these capabilities, but the bottom-up construction of artificial structures with the dimensions and complexity of viruses and other subcellular components remains challenging. Here we show that natural assembly principles can be combined with the methods of DNA origami(15-24) to produce gigadalton-scale structures with controlled sizes. DNA sequence information is used to encode the shapes of individual DNA origami building blocks, and the geometry and details of the interactions between these building blocks then control their copy numbers, positions and orientations within higher-order assemblies. We illustrate this strategy by creating planar rings of up to 350 nanometres in diameter and with atomic masses of up to 330 megadaltons, micrometre-long, thick tubes commensurate in size to some bacilli, and three-dimensional polyhedral assemblies with sizes of up to 1.2 gigadaltons and 450 nanometres in diameter. We achieve efficient assembly, with yields of up to 90 per cent, by using building blocks with validated structure and sufficient rigidity, and an accurate design with interaction motifs that ensure that hierarchical assembly is self-limiting and able to proceed in equilibrium to allow for error correction. We expect that our method, which enables the self-assembly of structures with sizes approaching that of viruses and cellular organelles, can readily be used to create a range of other complex structures with well defined sizes, by exploiting the modularity and high degree of addressability of the DNA origami building blocks used.