PURPOSE: Caffeine improves cycling time trial (TT) performance; however, it is unknown whether caffeine is ergogenic when competing against other riders. The aim of this study was to investigate whether caffeine improves performance during a 4-km cycling TT when riding against a virtual opponent, and whether it is associated with increased muscle activation and at the expense of greater end-exercise central and peripheral fatigue. METHODS: Using a randomized, crossover, and double-blind design, eleven well-trained cyclists completed a 4-km cycling TT alone without supplementation (CON), or against a virtual opponent after ingestion of placebo (OP-PLA) or caffeine (5 mg RESULTS: Compared to CON, OP-PLA improved 4-km cycling TT performance (P = 0.018), and OP-CAF further improved performance when compared to OP-PLA (P = 0.050). Muscle activation was higher in OP-PLA and OP-CAF than in CON throughout the trial (P = 0.003). The pre- to post-exercise reductions in voluntary activation and potentiated twitch force were, however, similar between experimental conditions (P > 0.05). Compared to CON, OP-PLA increased the rating of perceived exertion during the first 2 km, but caffeine blunted this increase with no difference between the OP-CAF and CON conditions. CONCLUSIONS: Caffeine is ergogenic when riding against a virtual opponent, but this is not due to greater muscle activation or at the expense of greater end-exercise central or peripheral fatigue.