Purpose: To compare game activity demands between female and male semi-professional basketball players. Methods: Female (n = 12) and male (n = 12) semi-professional basketball players were monitored across three competitive games. Time-motion analysis procedures quantified player activity into pre-defined movement categories across backcourt (BC) and frontcourt (FC) positions. Activity frequencies, durations, and distances were calculated relative to live playing time (∙min-1). Work:rest ratios were also calculated using the video data. Game activity was compared between genders for each playing position and all players. Results: Female players performed at greater running work-rates than males (45.7 ± 1.4 vs. 42.1 ± 1.7 m∙min-1, P = 0.05), while male players performed more dribbling than females (2.5 ± 0.3 vs. 3.0 ± 0.2 s∙min-1; 8.4 ± 0.3 vs. 9.7 ± 0.7 m∙min-1, P = 0.05). Positional analyses revealed female BC players performed more low-intensity shuffling (P = 0.04) and jumping (P = 0.05), as well as longer (P = 0.04) jogging durations than male BC players. Female FC players executed more upper-body activity (P = 0.03) and larger work:rest ratios (P < 0.001) than male FC players. No significant gender differences were observed in the overall intermittent demands, distance traveled, high-intensity shuffling activity, and sprinting requirements during game-play. Conclusions: These findings highlight that gender-specific running and dribbling differences might exist in semi-professional basketball. Further, position-specific variations between female and male basketball players should be considered. These data may prove useful in the development of gender-specific conditioning plans relative to playing position in basketball.