Backward walking is used increasingly as a rehabilitation exercise for stroke and diabetic peripheral neuropathy patients to improve strength and balance. However, it is unclear how visual referencing affects backward and forward walking. In this study, we evaluated spatiotemporal gait characteristics changes due to visual referencing while backward/forward walking. Sixteen healthy young participants were recruited in this study. All participants walked for 2 min with and without visual referencing in the virtual reality environment. While walking backward participants faced the virtual reality screen similar to forward walking, but their treadmill belt direction of movement was reversed. All participants walked at their preferred speed. We found that backward walking with visual reference affected symmetry in step length (p < 0.05) and step width (p < 0.001). Backward walking increased variability in step length (p < 0.001) and COM side excursions (p < 0.01) but also increased base of support through increased step width (p < 0.02). We also found backward walking with visual reference had significantly increased double support time (p < 0.001) and reduced swing time (p < 0.001). We also found that backward walking does not predispose to slip and trip risk, thereby, reduced foot contact velocity (p < 0.0001) and increased foot clearance (p < 0.0001). The findings of this study will help understand the effects of visual reference in backward and forward walking enables clinicians to design patient-centered rehabilitation exercises. © 2020 Elsevier B.V.