The purpose was to investigate the effects of CYP1A2 -163C > A polymorphism on the effects of acute caffeine (CAF) supplementation on anaerobic power in trained males. Sixteen trained males (age: 21.6 ± 7.1 years; height: 179.7 ± 5.6 cm; body mass: 72.15 ± 6.8 kg) participated in a randomized, double-blind, placebo (PLA) controlled crossover design. Participants supplemented with CAF (6 mg/kg of body mass) and an isovolumetric PLA (maltodextrin) in random order and separated by 7 days, before an all-out 30-s anaerobic cycling test to determine peak, average, and minimum power output, and fatigue index. Genomic deoxyribonucleic acid was extracted to identify each participants CYP1A2 genotype. Six participants expressed AA homozygote and 10 expressed C alleles. There was a treatment by genotype interaction for peak power output (p = .041, η2 = .265, observed power = 0.552) with only those expressing AA genotype showing improvement following CAF supplementation compared with PLA (CAF: 693 ± 108 watts vs. PLA: 655 ± 97 watts; p = .039), while no difference between treatments was noted in those expressing C alleles (CAF: 614 ± 92 watts vs. PLA: 659 ± 144 watts; p = .135). There were no other interaction or main effects for average or minimum power output, or fatigue index (p > .05). In conclusion, the ingestion of 6 mg/kg of CAF improved peak power output only in participants with the AA genotype compared with PLA; however, expression of the CYP1A2 did not influence average or minimum power output or fatigue index.