Verbal route descriptions are common in our daily lives that give us wayfinding directions. They also are important in cognitive research as they lend insight on processes associated with wayfinding. This paper reports a study that investigates the influence of familiarity and spatial abilities on acquiring spatial knowledge from verbal route directions. The familiarity of the participant was removed by replacing all names of spatial entities in the route instructions given by the same person. Specifically, the types of acquired spatial knowledge addressed are direction, distance, and configurational aspects of sketched maps. Results show that familiarity plays a crucial role on acquisition of spatial knowledge at the survey level. In particular, familiarity leads to fewer errors in directional estimation, but overestimation of distance. Spatial abilities further influence one’s knowledge of distance such that higher spatial abilities lead to more accurate distance estimation in new environments. With that said, lower spatial abilities do not contribute to distance estimation in both familiar and new environments. Furthermore, measures on sketch maps show that familiarity does not lead to dramatically different sketch maps while variation exist. These results also point out the necessity of follow-up studies to address the orientation specificity in familiar and unfamiliar environment.