Dongsheng Lei1 Alexander Marras2 Jianfang Liu1 Chao-Min Huang2 Lifeng Zhou2 Carlos Castro2 Hai-Jun Su2 Gang Ren1

1, Lawrence Berkeley National Laboratory, Berkeley, California, United States
2, The Ohio State University, Columbus, Ohio, United States

Scaffolded DNA origami has proven to be a powerful and efficient technique to fabricate functional nanomachines by programming the folding of a single-stranded DNA viral genome into three-dimensional (3D) nanostructures, designed to be precisely motion-controlled. Although two-dimensional (2D) imaging of DNA nanomachines using transmission electron microscopy and atomic force microscopy suggested these nanomachines are dynamic in 3D, geometric analysis based on 2D imaging was insufficient to uncover the exact motion in 3D. Here we use individual-particle electron tomography method and reconstruct 129 density maps from 129 individual DNA origami Bennett linkage mechanisms at ~6-14 nm resolution. The statistical analyses of these conformations lead to understanding the 3D structural dynamics of Bennett linkage mechanisms. Moreover, our effort provides, for the first-time, experimental verification of the theoretical kinematics model of DNA origami, which can be used as feedback to improve the design and control of motion via optimized DNA sequences.