The objectives of this study were to determine symmetry and sex differences in knee kinematics and anterior cruciate ligament (ACL) elongation waveforms in healthy athletes without a history of a knee injury during fast running, drop jump, and 180° internal/external rotation hops. It was hypothesized that knee abduction angle and ACL relative elongation would be greater in women than in men during all activities. Bilateral knee kinematics and ACL relative elongation were determined in 19 collegiate athletes using dynamic biplane radiography. Sex differences in kinematics and ACL relative elongation waveforms were identified using statistical parametric mapping. Average absolute side-to-side differences (SSD ) in kinematics and ACL relative elongation waveforms were determined for each activity. Women had up to 2.3° (all p < 0.05) less knee adduction angle and had greater ACL relative elongation (max. 4.8%-9.2%; all p < 0.01) than men during all activities, in support of the hypotheses. SSD in kinematics were 1.4 mm and 5.5° or less in all components of translation and rotation, respectively, while SSD in ACL relative elongation was 3.6% or less across all activities. Greater ACL relative elongation across a variety of activities may make women more susceptible to ACL injury than men. This study provides valuable reference data for identifying abnormal asymmetry in knee kinematics and ACL elongation in athletes after the ACL injury. These novel results improve our understanding of ACL elongation during demanding athletic activities and may help guide the development of sex-specific risk screening metrics, return to play assessments, and rehabilitation protocols after the ACL injury.