Sprecher
Beschreibung
The goal of this project is to develop a synthetic system that can sense, process and actuate molecular information in a programmable manner. For this purpose, we developed a DNA based reaction cascade that undergoes two steps, generating two temporally resolved output signals. The two reaction centers, A and B, are localized few nanometers apart inside a DNA origami compartment. Upon addition, the trigger DNA strand interacts with center A liberating an intermediate species that sequentially reacts with center B. Recording the first and second step of the reaction cascade by FRET, we investigate whether and how the spatial arrangement of the input species, different types and concentrations of crowding agents, as well as the attachment of different cargo sizes to the trigger strand, affect the kinetic signature of the process; both at the ensemble and single-molecule levels. We also show that the output signal can be coupled to a chain growth event, leading to the formation of filament-like appendices on one side of the origami cage. Such a large-scale morphological change is hypothesized to alter the overall hydrodynamic properties of the nano-object and affect its random diffusion behavior in solution. Hence, our model system enables not only monitoring and analyzing signal transmission within a nanosized environment but permits also to couple an input signal to a physical observable. We envision that the programmability of the DNA origami object, both inside and outside the cavity, will offer a versatile tool for studying reaction kinetics at the nanoscale and creating more sophisticated nano-sized objects that can self-propel in response to certain stimuli.
