Harnessing a paper-folding mechanism for reconfigurable DNA origami.

The paper-folding mechanism has been widely adopted in building of reconfigurable macroscale systems because of its unique capabilities and advantages in programming variable shapes and stiffness into a structure1-5. However, it has barely been exploited in the construction of molecular-level systems owing to the lack of a suitable design principle, even though various dynamic structures based on DNA self-assembly6-9 have been developed10-23. Here we propose a method to harness the paper-folding mechanism to create reconfigurable DNA origami structures. The main idea is to build a reference, planar wireframe structure24 whose edges follow a crease pattern in paper folding so that it can be folded into various target shapes. We realized several paper-like folding and unfolding patterns using DNA strand displacement25 with high yield. Orthogonal folding, repeatable folding and unfolding, folding-based microRNA detection and fluorescence signal control were demonstrated. Stimuli-responsive folding and unfolding triggered by pH or light-source change were also possible. Moreover, by employing hierarchical assembly26 we could expand the design space and complexity of the paper-folding mechanism in a highly programmable manner. Because of its high programmability and scalability, we expect that the proposed paper-folding-based reconfiguration method will advance the development of complex molecular systems.

Predicting the Free-Form Shape of Structured DNA Assemblies from Their Lattice-Based Design Blueprint.

Structured DNA assemblies have been designed primarily on a three-dimensional lattice because it is easy to arrange and cross-link the helices there. However, when we design free-form structures including wireframes and topologically closed circular objects on a lattice, artificially stretched bonds connecting bases are inevitably and arbitrarily formed. They often lead to nonconvergence or convergence to a wrong configuration in computational analysis to predict the equilibrium shape of the structure when started from its lattice-based configuration, which hinders the design process of free-form structures. Here, we present a computational procedure enabling the shape prediction of free-form structures from their lattice-based design blueprint without any convergence issue. It automatically partitions the structure into substructures and relocates them into a new configuration. When the analysis for calculating the equilibrium shape begins from this configuration, no convergence issue occurs because substructures and stretched bonds connecting them do not overlap and intertwine each other during analysis. Using the proposed approach, we could obtain the free-form shape of a comprehensive set of wireframe and circular structures accurately and quickly. We further demonstrated that it also facilitated a design of wireframe structures with nonstraight edges.

The flexibility-based modulation of DNA nanostar phase separation.

Phase separation of biomolecules plays key roles in physiological compartmentalization as well as pathological aggregation. A deeper understanding of biomolecular phase separation requires dissection of a relation between intermolecular interactions and resulting phase behaviors. DNA nanostars, multivalent DNA assemblies of which sticky ends define attractive interactions, represent an ideal system to probe this fundamental relation governing phase separation processes. Here, we use DNA nanostars to systematically study how structural flexibility exhibited by interacting species impacts their phase behaviors. We design multiple nanostars with a varying degree of flexibility using single-stranded gaps of different lengths in the arm of each nanostar unit. We find that structural flexibility drastically alters the phase diagram of DNA nanostars in such a way that the phase separation of more flexible structures is strongly inhibited. This result is not due to self-inhibition from the loss of valency but rather ascribed to a generic flexibility-driven change in the thermodynamics of the system. Our work provides not only potential regulatory mechanisms cells may exploit to dynamically control intracellular phase separation but also a route to build synthetic systems of which assembly can be controlled in a signal dependent manner.

Rapid Computational Analysis of DNA Origami Assemblies at Near-Atomic Resolution.

Structural DNA nanotechnology plays an ever-increasing role in advanced biomolecular applications. Here, we present a computational method to analyze structured DNA assemblies rapidly at near-atomic resolution. Both high computational efficiency and molecular-level accuracy are achieved by developing a multiscale analysis framework. The sequence-dependent relative geometry and mechanical properties of DNA motifs are characterized by the all-atom molecular dynamics simulation and incorporated into the structural finite element model successfully without significant loss of atomic information. The proposed method can predict the three-dimensional shape, equilibrium dynamic properties, and mechanical rigidities of monomeric to hierarchically assembled DNA structures at near-atomic resolution without adjusting any model parameters. The calculation takes less than only 15 min for most origami-scale DNA nanostructures consisting of 7000-8000 base-pairs. Hence, it is expected to be highly utilized in an iterative design-analysis-revision process for structured DNA assemblies.

Design Approaches and Computational Tools for DNA Nanostructures.

Designing a structure in nanoscale with desired shape and properties has been enabled by structural DNA nanotechnology. Design strategies in this research field have evolved to interpret various aspects of increasingly more complex nanoscale assembly and to realize molecular-level functionality by exploring static to dynamic characteristics of the target structure. Computational tools have naturally been of significant interest as they are essential to achieve a fine control over both shape and physicochemical properties of the structure. Here, we review the basic design principles of structural DNA nanotechnology together with its computational analysis and design tools.

Configurational Design of Mechanical Perturbation for Fine Control of Twisted DNA Origami Structures.

DNA origami nanotechnology allows us to rationally design molecular devices with arbitrary shapes and properties through programming the sequence of DNA bases for their directed self-assembly. Despite its remarkable shape programmability, it has not been fully explored yet how to precisely control the twisted shape of DNA origami structures shown to be important in controlling the physical properties of DNA devices, building DNA superstructures, and synthesizing macroscopic soft materials with targeted properties. Here, we demonstrate that designing the spatial configuration of mechanical strain energies induced by base pair (BP) insertions and deletions can effectively modulate the twist rate of DNA origami structures with a fine resolution. To illustrate, various six-helix bundles (6HB) were successfully constructed whose twist rate was precisely tuned with a mean increment of 1.8° per 21-BP-long unit block. We also show that locally relaxing the strain energy via engineered gaps, short unpaired nucleotides (NTs), can widen the range of achievable twist rate with fine controllability. The proposed configurational design approach is expected to expand the feasible design space of twisted DNA origami structures for their various potential applications with target functionalities.

Investigating the sequence-dependent mechanical properties of DNA nicks for applications in twisted DNA nanostructure design.

DNA nick can be used as a design motif in programming the shape and reconfigurable deformation of synthetic DNA nanostructures, but its mechanical properties have rarely been systematically characterized at the level of base sequences. Here, we investigated sequence-dependent mechanical properties of DNA nicks through molecular dynamics simulation for a comprehensive set of distinct DNA oligomers constructed using all possible base-pair steps with and without a nick. We found that torsional rigidity was reduced by 28-82% at the nick depending on its sequence and location although bending and stretching rigidities remained similar to those of regular base-pair steps. No significant effect of a nick on mechanically coupled deformation such as the twist-stretch coupling was observed. These results suggest that the primary structural role of nick is the relaxation of torsional constraint by backbones known to be responsible for relatively high torsional rigidity of DNA. Moreover, we experimentally demonstrated the usefulness of quantified nick properties in self-assembling DNA nanostructure design by constructing twisted DNA origami structures to show that sequence design of nicks successfully controls the twist angle of structures. Our study illustrates the importance as well as the opportunities of considering sequence-dependent properties in structural DNA nanotechnology.