Analysis of the flexibility profile of basketball players (BPs) can reveal differences in range of motion (ROM) by gender and also identify those players who are at higher risk for sports injuries. A descriptive observational study was conducted to determine the lower extremity flexibility profile of sixty-four basketball players and gender differences to identify players at higher risk of injury due to limited and asymmetric ROM in one or more movements. Participants: Sixty-four (33 male and 31 female) competitive athletes from the national leagues of the Spanish basketball league system participated in the present study (power of sample size ≥0.99). The eight passive ROM tests of the hip, knee and ankle were assessed using the ROM-SPORT battery. Each player completed a questionnaire on age, basic anthropometric data, dominant extremities, and training and sport-related variables. The lower extremity flexibility profile was established at 15° and 10° hip extension (HE), 39° and 38° ankle dorsiflexion with knee extended (ADF-KE), 40° and 39° ankle dorsiflexion with knee flexed (ADF-KF), 43° and 43° hip abduction (HAB), 75° and 61° hip abduction with the hip flexed (HAB-HF), 78° and 83° hip flexion with the knee extended (HF-KE), 134° and 120° knee flexion (KF), and 145° and 144° hip flexion (HF) by male and female basketball players, respectively. Sex differences in HE, HAB-HF, and KF were observed in BPs (p ≤ 0.01; Hedges’ g ≥ 1.04). Players reported limited ROM in ADF-KF, HE, HAB-HF, HF-KE, and KF; and asymmetric ROM mainly in HE, ADF-KE, KF, ADF-KF, and HF-KE. In conclusion, this study provides gender-specific lower extremity flexibility profile scores in BPs that can help athletic trainers and athletic and conditioning trainers to identify those players who are at higher risk of injury due to abnormal ROM scores.