The need for regional planning is increasingly important for effective Florida panther (Puma concolor coryi) (Bangs 1898) conservation and is essential for protecting enough habitat in South Florida to ensure a viable population. We used two decades of radio telemetry data and geographic information system (GIS) software to develop a regional blueprint for landscape restoration that enhances dispersal, facilitates population colonization, and could be the basis for future land use decisions in the range of the endangered Florida panther. We identified 923,576 ha of forests in an 18-county study area that is a barrier-rich patchwork of land uses. A least cost path analysis simulated natural colonization events and can be used to identify landscape linkages and conservation networks for the panther. Our analysis of planned development permits suggests that large-scale land protection must happen quickly. The alternatives are managing an isolated, heavily managed population or large-scale landscape restoration that is probably economically unfeasible.