Modular reconfiguration of DNA origami assemblies using tile displacement.

The power of natural evolution lies in the adaptability of biological organisms but is constrained by the time scale of genetics and reproduction. Engineeringartificial molecular machines should not only include adaptability as a core feature but also apply it within a larger design space and at a faster time scale. A lesson from engineering electromechanical robots is that modular robots can perform diverse functions through self-reconfiguration, a large-scale form of adaptation. Molecular machines made of modular, reconfigurable components may form the basis for dynamic self-reprogramming in future synthetic cells. To achieve modular reconfiguration in DNA origami assemblies, we previously developed a tile displacement mechanism in which an invader tile replaces another tile in an array with controlled kinetics. Here, we establish design principles for simultaneous reconfigurations in tile assemblies using complex invaders with distinct shapes. We present toehold and branch migration domain configurations that expand the design space of tile displacement reactions by two orders of magnitude. We demonstrate the construction of multitile invaders with fixed and variable sizes and controlled size distributions. We investigate the growth of three-dimensional (3D) barrel structures with variable cross sections and introduce a mechanism for reconfiguring them into 2D structures. Last, we show an example of a sword-shaped assembly transforming into a snake-shaped assembly, illustrating two independent tile displacement reactions occurring concurrently with minimum cross-talk. This work serves as a proof of concept that tile displacement could be a fundamental mechanism for modular reconfiguration robust to temperature and tile concentration.

Digital nanoreactors to control absolute stoichiometry and spatiotemporal behavior of DNA receptors within lipid bilayers.

Interactions between membrane proteins are essential for cell survival but are often poorly understood. Even the biologically functional ratio of components within a multi-subunit membrane complex-the native stoichiometry-is difficult to establish. Here we demonstrate digital nanoreactors that can control interactions between lipid-bound molecular receptors along three key dimensions: stoichiometric, spatial, and temporal. Each nanoreactor is based on a DNA origami ring, which both templates the synthesis of a liposome and provides tethering sites for DNA-based receptors (modelling membrane proteins). Receptors are released into the liposomal membrane using strand displacement and a DNA logic gate measures receptor heterodimer formation. High-efficiency tethering of receptors enables the kinetics of receptors in 1:1 and 2:2 absolute stoichiometries to be observed by bulk fluorescence, which in principle is generalizable to any ratio. Similar single-molecule-in-bulk experiments using DNA-linked membrane proteins could determine native stoichiometry and the kinetics of membrane protein interactions for applications ranging from signalling research to drug discovery.

DNA Strand-Displacement Temporal Logic Circuits.

Molecular circuits capable of processing temporal information are essential for complex decision making in response to both the presence and history of a molecular environment. A particular type of temporal information that has been recognized to be important is the relative timing of signals. Here we demonstrate the strategy of temporal memory combined with logic computation in DNA strand-displacement circuits capable of making decisions based on specific combinations of inputs as well as their relative timing. The circuit encodes the timing information on inputs in a set of memory strands, which allows for the construction of logic gates that act on current and historical signals. We show that mismatches can be employed to reduce the complexity of circuit design and that shortening specific toeholds can be useful for improving the robustness of circuit behavior. We also show that a detailed model can provide critical insights for guiding certain aspects of experimental investigations that an abstract model cannot. We envision that the design principles explored in this study can be generalized to more complex temporal logic circuits and incorporated into other types of circuit architectures, including DNA-based neural networks, enabling the implementation of timing-dependent learning rules and opening up new opportunities for embedding intelligent behaviors into artificial molecular machines.

A Loser-Take-All DNA Circuit.

DNA-based neural networks are a type of DNA circuit capable of molecular pattern recognition tasks. Winner-take-all DNA networks have been developed to scale up the complexity of molecular pattern recognition with a simple molecular implementation. This simplicity was achieved by replacing negative weights in individual neurons with lateral inhibition and competition across neurons, eliminating the need for dual-rail representation. Here we introduce a new type of DNA circuit that is called loser-take-all: an output signal is ON if and only if the corresponding input has the smallest analog value among all inputs. We develop a DNA strand-displacement implementation of loser-take-all circuits that is cascadable without dual-rail representation, maintaining the simplicity desired for scalability. We characterize the impact of effective signal concentrations and reaction rates on the circuit performance, and derive solutions for compensating undesired signal loss and rate differences. Using these approaches, we successfully demonstrate a three-input loser-take-all circuit with nine unique input combinations. Complementary to winner-take-all, loser-take-all DNA circuits could be used for recognition of molecular patterns based on their least similarities to a set of memories, allowing classification decisions for patterns that are extremely noisy. Moreover, the design principle of loser-take-all could be more generally applied in other DNA circuit implementations including k-winner-take-all.

Capturing Plastic Surgery on Film-Making Reconstruction Visible.

The Swiss Plastic Surgery Association (https://plasticsurgery.ch/en/) decided to produce a corporate video to illustrate the concept of "plastic surgery of confidence" to the public. We show the diversity of specializations and the vast range of tasks that surgeons passionately handle day in and day out. We wanted to convey 2 main messages: first, that plastic surgery is more than just cosmetic surgery, and second, that plastic surgery in Switzerland is synonymous with quality and confidence. We selected 17 topics that we felt had good filmic potential and would best explain to the public what plastic surgery is about. This included the selection of appropriate patients, experts, and locations from all over the country. We thought it crucial to show the initial preoperative situation, as only in this case would the achievement of reconstruction be evident and comprehensive to the layman audience. The actual production was filmed in 5 different locations and took 5 days of shooting. We recorded 17 surgeons, 9 patients, and about 30 voluntary background actors. From 23 hours of footage, we created a 7 minute, 22 second corporate video, recorded in 3 of the Swiss national languages. The video was presented to the public online in June 2016, on the same day as the National Open Day of Plastic Surgery in Switzerland. The video is available online. We evaluated the impact of the video using a questionnaire for lay people and observed that it could substantially improve the perception of our specialty, especially regarding the reconstructive aspect. We feel that a freely available corporate video is a very useful means to promote plastic surgery and help patients better understand what it is all about.