molecule via a transition metal ion and subsequently closed by another crescentshaped molecule in a single cyclization reaction. In strategy B "entwining," two crescent-shaped molecules are linked together via a transition metal ion, followed by a double cyclization reaction to close them at once. In each case, the transition metal ion can be removed after serving its purpose, enabling the free relative movements of the two rings.Mechanically interlocked architectures are appealing building blocks for the realization of many functional devices. Importantly, they empower broad applications in nanomaterials, nanorobotics, and nanomachines. [18][19][20][21] For instance, catenanes have been exploited as switches, [22] rotary motors, [23,24] and sensors. [25][26][27][28][29][30] Rotaxanes have been used as molecular shuttles, [31,32] switches in molecular electronics, [33] control of chemical synthesis, [34] and force-generating components for molecular elevators [35] and pumps. [36] An alternative pathway to create topologically complex molecular architectures is structural DNA nanotechnology, which exploits DNA as both genetic and construction material. [37][38][39][40] The first topological DNA structures were demonstrated by Seeman and co-workers, who synthesized a variety of DNA knots and Borromean rings. [41,42] It was then followed by the construction of DNA catenanes, [43][44][45][46] rotary motors, [47,48] a biohybrid nanoengine, [49] and logic circuits. [50] In 2010, DNA rotaxanes were reported. [51] Subsequently, chemically more rigid, [52] light-switchable, [53] and Daisy-chain rotaxanes, [54] DNA-nanoparticle rotaxanes [55] as well as catalytically active rotaxanes [56] were demonstrated. Despite being topologically well-defined, the as-fabricated DNA catenanes and rotaxanes were in general mechanically rather flexible and floppy, because they were made of single-or doublestranded DNA. By contrast, DNA origami-based interlocked architectures possess much better structural rigidity. The first attempt to realize such catenanes was carried out by Yan and co-workers, who followed an innovative scheme of molecular kirigami. [57] In 2016, Simmel and co-workers reported switchable DNA origami rotaxanes with long-range on-axis motion. [58] In this work, we experimentally demonstrate the hierarchical assembly of DNA origami catenanes templated by gold nanoparticles (AuNPs), taking inspirations from the transition metal-templated synthesis of catenanes developed by Sauvage and co-workers. DNA origami catenanes, which contain two, three or four interlocked rings are successfully created. In particular, the origami rings within the individual catenanes can be set free with respect to one another by releasing the interconnecting AuNPs through toehold-mediated strand displacement reactions.Mechanically interlocked molecules have marked a breakthrough in the field of topological chemistry and boosted the vigorous development of molecular machinery. As an archetypal example of the interlocked molecules, catenanes compri...