This species is found in a variety of habitats, ranging from relatively arid to mesic environments. They prefer wet areas, however, especially woodlands and thickets near streams and swamps. Also in suburban areas. The opportunistic denning and feeding habits of the Virginia opossum has led to the success of the species, especially in areas of habitat fragmentation. High reproductive potential further contributes to increasing population size (McManus 1974). Abandoned burrows, buildings, hollow logs, and tree cavities are generally used for den sites.

This species is found in Central America, from Costa Rica to Mexico and in the United States east of the Rocky Mountains, and north into southwestern Ontario, Canada (the northernmost locality reached by a marsupial). Some introduced populations are also found along the west coast of the United States and recently into British Columbia (Canada). Their range, limited by winter temperatures and snow depth, appears to be expanding northwards (Gardner 2005). This species can be found from lowlands to 3,000 m (Reid 1997).

There are no specific measures in place to protect the Virginia opposum, as it likely occurs in a number of protected areas throughout its range.