Kilwing Luzia, Lill Pascal, Nathwani Bhavik, Guerra Richard, Benson Erik, Liedl Tim, Shih William M
Faculty of Physics and Center for NanoScience (CeNS), Ludwig-Maximilians-University, Geschwister-Scholl-Platz 1, 80539 Munich, Germany.
Department of Cancer Biology, Dana-Farber Cancer Institute; Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115, United States.
ACS Nano. 2024 Jan 9;18(1):885-893. doi: 10.1021/acsnano.3c09522. Epub 2023 Dec 18.
DNA origami is a popular nanofabrication strategy that employs self-assembly of a long single scaffold strand, typically less than 10 kilobases in length, with hundreds of shorter staple strands into a desired shape. In particular, origami arranged as a single-layer rectangle has proven popular as flat pegboards that can display functionalities at staple-strand breakpoints, off the sides of the constituent double helices, with a ∼5.3 nm rhombic-lattice spacing. For applications that demand tighter spacing, functionalities can be displayed instead on the termini of helices of multilayer DNA origami. However, pegboards with the greatest addressable surface area are often found to be the most versatile. Given the practical limitations of the length of the scaffold that can be easily realized, designs that minimize the length of each helix would have advantages for maximizing the number of helices and therefore the number of addressable pixels on each terminal surface. Here we present an architecture for multilayer DNA origami displaying flush terminal interfaces from over 200 helices that each are only 5.3 turns in length. We characterize an example using cryo-EM imaging paired with single-particle analysis for further analysis of the global structure.
DNA折纸术是一种流行的纳米制造策略，它利用一条长的单支架链（通常长度小于10千碱基）与数百条较短的钉状链自组装成所需形状。特别地，排列成单层矩形的折纸术已被证明很受欢迎，它就像扁平的钉板一样，可以在钉状链断点处、在组成双螺旋的侧面之外展示功能，其菱形晶格间距约为5.3纳米。对于需要更紧密间距的应用，可以在多层DNA折纸术的螺旋末端展示功能。然而，具有最大可寻址表面积的钉板通常被发现是最通用的。考虑到易于实现的支架长度的实际限制，最小化每个螺旋长度的设计对于最大化螺旋数量以及因此每个末端表面上的可寻址像素数量具有优势。在这里，我们展示了一种多层DNA折纸术的架构，它展示了来自200多个螺旋的齐平末端界面，每个螺旋的长度仅为5.3圈。我们使用低温电子显微镜成像与单颗粒分析相结合的方法对一个示例进行了表征，以便对整体结构进行进一步分析。
