In spite of being a promising therapeutic modality, gene therapy has limited clinical applications, mostly due to the lack of spatiotemporal resolution and inadequate efficacy. Herein, we present a facile strategy to remotely control intracellular gene expression by using gold nanorod- (Au NR) derived, host-guest interaction-mediated supramolecular vesicles as a gene carrier and photothermal transducer. Upon pulsed laser irradiation, mild photothermal conditions dissociate supramolecular vesicles to release gene and simultaneously activate heat shock protein-70 promoter (Hsp70) for spatiotemporally initiating gene expression inside cancer cells. Subsequently, upon introducing a polymeric guest species specifically into cancer cells, the dissociated Au NRs functionalized with macrocyclic host molecules could re-aggregate rapidly in cells to retard exocytosis of these NRs, thereby allowing deferred photothermal therapy to enhance the overall therapeutic outcome.