Purpose: Asymmetric gait patterns are known to persist in patients with unilateral total hip arthroplasty (THA). Little is known about the effect of gender on gait asymmetry. This study aimed to determine whether gait asymmetry differs between genders in unilateral THA patients and can be explained by component positioning. Methods: 23 female and 10 male unilateral THA patients were included. Asymmetric hip kinematics during gait and component positioning were quantified using a combined computed tomography-based modelling and dual fluoroscopic imaging system. Multiple regression was performed to examine the unique and mediated contribution of gender to observed gait asymmetry. Results: Female and male patients differed most significantly in frontal plane gait asymmetry (p = 0.001) and bilateral difference of vertical femoral offset (p = 0.048). Compared with the native hip, the implanted hip exhibited significantly increased adduction (p = 0.007) in females but significantly increased abduction (p = 0.001) in males. The stem head of the implanted hip was more superiorly positioned compared to the native femoral head in female but more inferiorly positioned in male. Gender accounted for 41.5% variance of frontal plane gait asymmetry and was partially mediated by the bilateral difference of vertical femoral offset. Conclusions: Female unilateral THA patients exhibited significantly increased adduction in the implanted hip compared to the native hip, which is partially attributable to a more superiorly positioned femoral stem centre of rotation. The understanding of gender-specific differences of kinematic patterns may benefit female patients through targeted preoperative planning and postoperative rehabilitation.