Spatial navigation, the ability to explore, learn, and remember one’s environment, is a skill that is fundamental to everyday goals. Navigation is challenging to study because it requires understanding and integrating multiple sources of information from the environment and the body as well as many cognitive processes including perception, attention, memory, and decision making. Virtual Reality (VR), the presentation and experience of a synthetic sensory environment, provides a method to study spatial navigation in controlled, ecologically valid, and innovative ways. In this review, we describe VR as a tool to study spatial navigation with the perspective that basic science questions and applications mutually inform one another. We focus on how VR has advanced an understanding of the spatial cues, spatial knowledge, and individual differences involved in spatial navigation and discuss the role and implications of different VR technologies that allow for these investigations.