Evaluation of Structural Vulnerabilities in DNA Origami Nanoplatforms Using Ionizing Radiation

DNA origami holds great promise as a platform for diverse nanostructure assemblies, but its long‐term stability remains a key challenge, especially in application‐specific environments. In this study, we used two types of ionizing radiation to controllably induce strand breaks in three distinct DNA origami shapes, simulating long‐term damage and identifying structural weak points. Damage was assessed using atomic force microscopy (AFM), agarose gel electrophoresis (AGE), and melting point analysis. AFM revealed that overall structural integrity was retained even after exposure to extremely high radiation doses. However, melting point measurements showed a reduction in thermal stability, indicating an increased number of strand breaks, supported by AGE analysis of denatured samples. Vulnerabilities were most pronounced at exposed edges and single‐stranded regions, which are more accessible to reactive species generated during water radiolysis. This approach offers a simple method to identify DNA origami weak points and probe long‐term stability of DNA nanostructures in a fast and controlled way. The results also provide benchmarks for theoretical models and guidelines for the design of robust DNA origami for various applications.