“…Recently, DNA, as one of the most powerful materials and a moldable building block, has become very popular for constructing artificial architectures and molecular machines owing to the exquisite programmability, specificity and predictability of Watson–Crick base pairing. − To date, various DNA nanostructures and nanomachines, such as DNA origamis, tetrahedron, gears, switches, robots, tweezers, and walkers, , have been developed, which exhibited outstanding applications in various research fields. Among them, DNA walker, a type of dynamic DNA device consisting of the drive motor, the walking track, and the walking strand, in which the walking strand can move progressively along confined walking tracks once the initial equilibrium was broken by the drive motor, has particularly aroused great interest from scientists. , Initial DNA walkers were designed to run on the predefined one-dimensional (1D) linear track and the two-dimensional (2D) planar track. − Compared with 1D and 2D DNA walker, the recently reported three-dimensional (3D) DNA walker showcases higher walking efficiency and better signal amplification performance due to the high surface-to-volume ratio and increased capacity of 3D spherical tracks. , At present, 3D DNA walker, as an ideal and powerful analytical tool, had been engineered and employed for the detection of DNA, RNA, , protein, bacteria , and metal ions .…”