Road user fatalities account for one of the leading causes of preventable death in Latin America with pedestrians and bicyclists at higher risk for more extensive injuries as compared to other road users. Despite these vulnerable road user (VRU) risks, encouraging individuals to walk and cycle is an important public health strategy for addressing the region's obesity epidemic through promoting regular physical activity via active transportation (AT). However, in order to promote AT as a viable source of physical activity, safety of the VRU must be considered. Interventions to improve VRU safety are critical but a current understanding of the impact of these interventions in the region is lacking. The purpose of this systematic review and metasummary is to describe the effectiveness of interventions that have been implemented in Latin America to improve pedestrian and bicyclist safety. A systematic literature search of public health, policy and engineering databases was completed using search terms generated through the PICO method. The PRISMA framework was used to screen articles for inclusion and analysis. Eight articles detailing nine interventions across four countries were included for final synthesis and organized according to the Three E’s Model of Injury Prevention (Education, Engineering and Enforcement). Interventions assessed outcomes related to VRU safety ranging from attitudes and behaviors to fatal injuries, with only enforcement-based interventions reporting on the latter. Evidence-based interventions to inform the design of programs to promote AT and VRU safety in the region are lacking and should be of urgent public health priority.