Assessing the potential for threatened species to persist and spread within fragmented landscapes requires the identification of core areas that can sustain resident populations and dispersal corridors that can link these core areas with isolated patches of remnant habitat. We developed a set of GIS tools, simulation methods, and network analysis procedures to assess potential landscape connectivity for the Delmarva fox squirrel (DFS; Sciurus niger cinereus), an endangered species inhabiting forested areas on the Delmarva Peninsula, USA. Information on the DFS's life history and dispersal characteristics, together with data on the composition and configuration of land cover on the peninsula, were used as input data for an individual‐based model to simulate dispersal patterns of millions of squirrels. Simulation results were then assessed using methods from graph theory, which quantifies habitat attributes associated with local and global connectivity. Several bottlenecks to dispersal were identified that were not apparent from simple distance‐based metrics, highlighting specific locations for landscape conservation, restoration, and/or squirrel translocations. Our approach links simulation models, network analysis, and available field data in an efficient and general manner, making these methods useful and appropriate for assessing the movement dynamics of threatened species within landscapes being altered by human and natural disturbances.