We evaluated the biological and socio-economic effects of statewide limitation of mule deer (Odocoileus hemionus) hunting licenses, which began in Colorado in 1999. We implemented a before-after-control-impact (BACI) analysis of annual helicopter sex and age class surveys, collected as part of the Colorado Division of Wildlife's routine monitoring, to assess changes in adult male/adult female ratios and fawn/adult female ratios in response to this change in harvest management. Following statewide limitation and reduction of license sales (1999–2006), we observed increases in adult male/adult female ratios of 7.39 (SE = 2.36) to 15.23 (SE = 1.22) adult males per 100 adult females in moderately limited areas and of 17.55 (SE = 3.27) to 21.86 (SE = 2.31) adult males per 100 adult females in highly limited areas. We simultaneously observed reductions in fawn/adult female ratios in newly limited areas by as much as 6.96 (SE = 2.19) fawns per 100 females, whereas in areas that had previously been limited we observed stabilization of fawn/adult female ratios at levels lower than levels observed under the unlimited harvest management structure. An immediate decline of $7.86 million in annual revenue stemmed from the change in harvest management, but revenue subsequently rebounded. This study provides preliminary evidence of potential effects that other state and provincial wildlife management agencies may face as they consider shifting mule deer harvest management towards limited license scenarios.