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Single-Molecule Real-Time Visualization of DNA Unwinding by CMG Helicase
Published on: September 27, 2024
Single-Molecule Observation of the Photoregulated Conformational Dynamics of DNA Origami Nanoscissors
Elena M Willner1, Yuu Kamada2, Yuki Suzuki2,3
1Physik Department and Institute for Advanced Study, Technische Universität München, Am Coulombwall 4a, 85748, Garching near Munich, Germany.
Abstract:
We demonstrate direct observation of the dynamic opening and closing behavior of photocontrollable DNA origami nanoscissors using high-speed atomic force microscopy (HS-AFM). First the conformational change between the open and closed state controlled by adjustment of surrounding salt concentration could be directly observed during AFM scanning. Then light-responsive moieties were incorporated into the nanoscissors to control these structural changes by photoirradiation. Using photoswitchable DNA strands, we created a photoresponsive nanoscissors variant and were able to distinguish between the open and closed conformations after respective irradiation with ultraviolet (UV) and visible (Vis) light by gel electrophoresis and AFM imaging. Additionally, these reversible changes in shape during photoirradiation were directly visualized using HS-AFM. Moreover, four photoswitchable nanoscissors were assembled into a scissor-actuator-like higher-order object, the configuration of which could be controlled by the open and closed switching induced by irradiation with UV and Vis light.

