Purpose: This study aimed to investigate how rifle carriage and skiing speed during biathlon roller skiing affect range of motion (ROM) in joint angles and equipment (skis and poles), the vertical distance between shoulders and treadmill (vertdist), as well as possible sex differences associated with rifle carriage. Methods: Fourteen biathletes (6 women, 8 men) roller-skied on a treadmill at submaximal and simulated race speeds, with (WR) and without (NR) a rifle, using gears 3 and 2. Kinematical data for the whole body, poles, roller-skis, rifle, and treadmill were monitored using a 3D motion capture system. Movements determined as flexion/extension (x), abduction/adduction (y), and/or internal/external rotation (z) were analyzed for the hip, shoulder, thorax, knee, ankle, elbow, poles, and roller skis. ROM (the difference between maximal and minimal angles) in joints and equipment, and vertdist were analyzed over six skiing cycles during each condition (WR and NR) and speed. Results: The maximal vertdist was lower for WR compared with NR (gear 3: 1.53 ± 0.06 vs 1.54 ± 0.06 m; gear 2: 1.49 ± 0.06 vs 1.51 ± 0.06 m; both p < 0.001). ROM in the upper body was altered when roller skiing WR (movements decreased in thorax and shoulder (x) and increased in elbow (only gear 3) (x), thorax (only gear 2), and shoulder (y) and (z); all p < 0.05) and increased with speed, without differences between sexes (p > 0.05). Conclusion: Since rifle carriage and speed appear to affect the kinematics of roller skiing, coaches, and biathletes are advised to perform skiing technique training under competition-like conditions (i.e., at race speeds while carrying the rifle).