Clerks at the Department of Motor Vehicles (DMV) serve as gatekeepers in the quest to fill organ donor registries. Most people who join organ donor registries do so through their local driver's license bureau. To improve knowledge, attitudes, and behavior toward organ donation among DMV clerks, and to improve strategies for communicating with the public about organ donation, resulting in more people joining DMV-based organ donor registries. DMV offices in 8 counties in a southern state. A general survey of 1504 participants in 8 counties was conducted to assess knowledge, attitudes, beliefs, and behaviors toward organ donation. All 76 DMV clerks in those counties participated in surveys before and after a training intervention. DMV clerks received a 1-hour training intervention focused on the nature of the new organ donor registry, information about organ donation, and communication strategies for interacting with the public. Knowledge, attitudes, beliefs, behaviors, and behavioral intentions toward organ donation were measured for the general public survey and for DMV clerks. Clerks were more knowledgeable about brain death and religious stances on organ donation than the general public, but otherwise both groups shared similar knowledge, attitudes, beliefs, behaviors, and behavioral intentions toward organ donation. Overall the general public and clerks had favorable attitudes toward organ donation but low to moderate knowledge, beliefs, and behaviors and behavioral intentions. Training significantly increased DMV clerks' knowledge, attitude, beliefs, and behavioral intention toward donation. Organ donor registration rates were a mean of 14% higher in counties where clerks were trained than in control counties and were 9% higher than statewide. Training DMV clerks is an effective way to increase knowledge, attitudes, and beliefs for gatekeepers of organ donor registries and may increase donor registration rates among the public.