To determine whether monkeys can navigate a virtual environment, and whether they can represent a virtual environment in their brain, we trained two macaque monkeys to perform a navigation task in a virtual building and conducted behavioral analyses. After learning the task from the first starting point, they were trained to perform the navigation task from a second starting point which is already familiar to them during the first training session. The second training was completed faster than the first one. Furthermore, we found that the time required for a free-way-finding test from a novel starting point was shortened after the second training. These results suggest that monkeys can navigate a virtual environment and can construct a flexible representation of a virtual environment in their brain.