Transportation infrastructure projects have been planned and implemented faster than data can be collected to inform planning about the biodiversity safeguards that would be needed for these projects. There is a need for rapid, robust, and proactive large-scale approaches to identify species and regions most at risk. Although several large-scale studies have used systematic reviews and meta-analyses for some groups of species and regions, such studies have not been performed widely to inform road agencies about the impacts of roads on wildlife and how to better avoid or mitigate the negative impacts. Here, we have synthesized available data for large-scale studies and the main findings about species and regions most at risk. We have also identified current assessment needs and propose some guiding principles for more accurate large-scale assessments. We hope to contribute to raising awareness on how accurate large-scale assessments can help identify species and regions at risk due to roads and traffic and prioritize regions for assessments at the local scale for effective mitigation measures.