Virtual environments (VEs) provide an inexpensive way of conducting ecologically valid psychological research. The present study used a VE to demonstrate conditioned suppression, a behavioral model of anxiety, in a first-person perspective video game. During operant training, participants learned to shoot crates to find gold bars and thus score points in the game. Next, during Pavlovian conditioning, a colored light (i.e., conditioned stimulus: CS+) was followed by a white noise unconditioned stimulus (US) while a different colored light (CS−) was not paired with the US. Probe trials in a final testing phase were then used to assess suppression. We found significant suppression of accurate responding (shots hitting the designated targets) during the presence of the CS+ relative to the CS−, both in terms of total hits and hits as a proportion of total shots. Importantly, this effect emerged despite the overall level of operant responding being undiminished during the CS+. Our findings are consistent with related studies examining human behavior in real environments, and demonstrate the potential of VEs in combination with a modestly aversive CS to allow a detailed behavioral profile of anxiety to emerge.