How We Make DNA Origami
How We Make DNA Origami
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DOI：
10.1002/cbic.201700377
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发表时间：
2017-10-05
期刊：
影响因子：
3.2
通讯作者：
Dietz, Hendrik
中科院分区：
文献类型：
--
作者：
Wagenbauer, Klaus F.;Engelhardt, Floris A. S.;Dietz, Hendrik
DNA折纸自十年前发明以来吸引了大量关注，因为它似乎为创造定制的纳米级物体提供了无限的可能性。虽然DNA折纸的基本概念很容易理解，但在实际应用中使用定制DNA折纸需要详细的技术诀窍来设计和生产具有足够质量的颗粒，并在定制环境中以适当的浓度制备具有必要纯度的颗粒。对于该领域的新手来说，这种专有技术并不容易获得，因此减缓了DNA纳米技术领域以外的新应用出现的速度。为了促进更快的进展，我们在这篇文章中分享了我们近年来积累的制作和准备DNA折纸的经验。我们讨论了用于创建高级结构图案的设计解决方案，包括角和各种类型的铰链，这些设计方案扩展了更刚性的多层DNA折纸的设计空间，并提供了防止不必要的聚集和如何诱导多个DNA折纸构建块的特定齐聚的指南。此外，我们提供了详细的方案，并讨论了五种关键方法的预期结果，这些方法允许高效和无损地制备DNA折纸。这些方法包括琼脂糖凝胶纯化、分子截留膜过滤、聚乙二醇沉淀法、尺寸排除层析和超速离心沉淀法。我们在这里描述的创建高级设计主题的指南和详细的协议及其实验特征应该会为研究人员完成完整的DNA折纸生产工作流程降低障碍。
DNA origami has attracted substantial attention since its invention ten years ago, due to the seemingly infinite possibilities that it affords for creating customized nanoscale objects. Although the basic concept of DNA origami is easy to understand, using custom DNA origami in practical applications requires detailed know-how for designing and producing the particles with sufficient quality and for preparing them at appropriate concentrations with the necessary degree of purity in custom environments. Such know-how is not readily available for newcomers to the field, thus slowing down the rate at which new applications outside the field of DNA nanotechnology may emerge. To foster faster progress, we share in this article the experience in making and preparing DNA origami that we have accumulated over recent years. We discuss design solutions for creating advanced structural motifs including corners and various types of hinges that expand the design space for the more rigid multilayer DNA origami and provide guidelines for preventing undesired aggregation and on how to induce specific oligomerization of multiple DNA origami building blocks. In addition, we provide detailed protocols and discuss the expected results for five key methods that allow efficient and damage-free preparation of DNA origami. These methods are agarose-gel purification, filtration through molecular cut-off membranes, PEG precipitation, size-exclusion chromatography, and ultracentrifugation-based sedimentation. The guide for creating advanced design motifs and the detailed protocols with their experimental characterization that we describe here should lower the barrier for researchers to accomplish the full DNA origami production workflow.