“…Dynamic DNA nanotechnology with DNA as creating material has attracted much attention in the field of molecular and nanomachines due to the inherent molecular recognition property, programmable high-precision structure, and controllable motion of DNA. − A three-dimensional (3D) DNA walker is one of the most typical dynamic DNA nanomachines that concentrates DNA reaction strands on the surface of 3D particles (such as gold nanoparticles or silicon microparticles) and shows machine-like movements in response to external stimuli. − Owing to the excellent molecular recognition ability, self-driven capacity, accelerated reaction rate, and superior signal amplification capability, the 3D DNA walker has been applied in biosensing, intracellular imaging, molecular computation, and clinical diagnostics. − According to the anchoring type of walking arms, the 3D DNA walker can be divided into two categories: fixed-arm walker , and free-arm walker. , Nevertheless, these two types of walkers usually have to anchor the DNA tracks on the 3D landscapes via covalent or coordinate bonds, which complicates their construction processes. Moreover, the walkers have to at least statically anchor the tracks on the 3D landscapes, which hinders the relative motion between the tracks and arms, thereby reducing walking efficiency.…”