Sprecher
Beschreibung
Precise spatial control of proteins is critical for the construction of synthetic biological systems and the investigation of dynamic molecular processes. DNA origami offers a powerful platform for organizing biomolecules with nanometer precision; however, most existing reconfiguration strategies depend on thermal cycling, enzymatic reactions, or highly customized designs, which limit their reversibility, adaptability, and ease of implementation. Here, we present a robust isothermal transformation strategy that enables rapid, programmable rearrangement of protein patterns on shape-shifting DNA origami structures under constant ambient conditions (Figure 1). Starting from a stable DNA origami shape, we have shown in the past that adding a competitive set of staples can lead to isothermal transformation into a desired shape if a properly designed buffer is used, but this process is very slow, taking up to 42 days and involve defects-rich origamis1. Here, we show that, by systematically optimizing transformation conditions, we can achieve high-yield isothermal transformation between perfectly shaped origamis in less than six hours. The method is highly versatile: it supports a wide range of shape transformations, including between 2D and 3D morphologies, multiple sequential transitions, and even reversible switching, all without requiring temperature changes, enzymatic assistance, or specialized origami designs. We covalently attach biotin-labeled DNA handles to the scaffold strand, allowing site-specific placement of proteins such as streptavidin. Their spatial organization can then be precisely modulated through the addition of reprogrammed staple sets corresponding to distinct DNA origami geometries. This enzyme-free, design-flexible approach provides a generalizable framework for reconfigurable protein patterning, offering new way for constructing adaptive nanodevices and responsive molecular architectures under biologically relevant, isothermal conditions.
Reference
1. Rossi-Gendron, C., El Fakih, F., Bourdon, L. et al. Isothermal self-assembly of multicomponent and evolutive DNA nanostructures. Nat. Nanotechnol. 18, 1311–1318 (2023). https://doi.org/10.1038/s41565-023-01468-2
