The brushtail possum (Trichosurus vulpecula) is in decline throughout much of its natural range with the notable exception of urban areas and a few islands. In some urban areas, its density can be high enough to cause conflict with householders. We provide the first genetic-based study of dispersal for T. vulpecula in the urban environment. Seven microsatellite markers were used to investigate genetic structure of adult male (n = 53) and female (n = 39) possums sampled from mainland suburbs of Sydney, and on nearby Scotland Island. Samples from Scotland Island also provide an opportunity to assess the effect of isolation on genetic variability, which, as theory would predict, was significantly lower than observed in mainland samples. Male-biased dispersal was inferred from patterns of relatedness between individuals of each sex. Average relatedness was significantly higher between adult female possums than between adult male possums sampled within the same garden area. In addition, males were genotypically more similar to one another at substantially greater geographic distances than females. Along with male-biased dispersal, strong localised genetic structure for both sexes infers generally high philopatry. Dispersal distances were greatest for adult male possums sampled from mainland locations. However, even for these males, genotypic similarity between possums separated by distances further than ∼900 m was lower than the sample average, suggesting infrequent dispersal beyond this distance. Knowledge of dispersal patterns is important to managing the density levels of overabundant T. vulpecula in urban areas.