Nano-impact electrochemistry (NIE) has made significant advances in the fields of electrochemistry and analytical chemistry. Nevertheless, the choice of entities used for NIE is usually confined to inorganic nanoparticles. The interest in DNA nanotechnology prompts the exploration of DNA materials as signal sources for NIE. In this study, tetrahedral DNA nanostructure (TDNA) is designed as an example. Various sizes and base compositions of TDNAs are assembled. Corresponding NIE signals are successfully detected at the interface of gold ultramicroelectrode. The results indicate that 3D DNA nanostructures can proficiently replicate the roles of inorganic nanoparticles in NIE without requiring additional modifications. Furthermore, it is demonstrated that TDNA not only adheres to the electrode surface, acting as a diffusion barrier but also exhibits NIE effects through base redox reactions and charge conduction. The findings supplement new types of materials with NIE effects and may inspire novel bioanalytical applications.