Abstract 2D transition metal dichalcogenides (TMDCs) as MoS 2 , possess intrinsic properties, such as ultra‐high surface‐to‐volume ratio, that make them ideal platforms in the construction of heterostructures for the loading and spatial confinement of functional components. A key feature of this kind of nanohybrids is the ability to control the reversibility of the assembly via interfacial engineering. In this study, the construction of a stimuli‐responsive nanohybrid based on DNA‐templated 2D heterostructures is reported. DNA strands are employed both as dynamic linkers on MoS 2 and nanoscale templates for the integration of Ag 2 S nanoparticles, which in turn imparted photothermal responsiveness to the heterostructure; this feature facilitated the controlled disassembly of the system. As proof of concept, various functional moieties—including a fluorophore, quantum dots, and an enzyme—are selectively tethered to the 2D scaffold via DNA conjugation and released from the MoS 2 surface in response to local heat induced by 808 nm light irradiation. This further allowed us to control the biocatalytic activity of cellulase in the hydrolysis of the biopolymer cellulose by phototriggering the enzyme release from MoS 2. In summary a stimuli‐responsive DNA‐templated mixed‐dimensional heterostructure is developed for the spatial confinement and phototriggered release of target molecules from a 2D nanomaterial platform.