Gaming industry employees are at higher risk than the general population for health conditions including gambling disorder. Responsible gambling training programs, which train employees about gambling and gambling-related problems, might be a point of intervention. However, such programs tend to use a "one-size-fits-all" approach rather than multiple tiers of instruction. We surveyed employees of one Las Vegas casino (n = 217) and one online gambling operator (n = 178) regarding their gambling-related knowledge and opinions prior to responsible gambling training, to examine the presence of natural knowledge groups among recently hired employees. Using k-means cluster analysis, we observed four natural groups within the Las Vegas casino sample and two natural groups within the online operator sample. We describe these natural groups in terms of opinion/knowledge differences as well as distributions of demographic/occupational characteristics. Gender and language spoken at home were correlates of cluster group membership among the sample of Las Vegas casino employees, but we did not identify demographic or occupational correlates of cluster group membership among the online gambling operator employees. Gambling operators should develop more sophisticated training programs that include instruction that targets different natural knowledge groups.