Introduction: Gait is a major function of independence that determines the quality of life, participation, and restriction. Gait assessment in physical therapy practice is fundamental for assessing musculoskeletal conditions. This study planned to determine the extent and barriers of using gait assessment tools by physical therapists in clinical practice in Saudi Arabia. Method: A cross-sectional design was used. A standardized survey was sent through e-mail and social media to physical therapists working in hospitals, clinics, and rehabilitation centers in different Saudi Arabian regions. Results: A total of 320 physical therapists from different regions in Saudi Arabia participated. There was a significant relationship between using gait assessment tools by physical therapists and patient groups (p = 0.002), receiving training (p = 0.001), dealing with patients who suffer from gait problems (p = 0.001), and visiting the gait laboratory (p = 0.001). Physical therapists’ use of gait assessment tools for musculoskeletal conditions in clinical practice in Saudi Arabia was limited. The primary barrier preventing physical therapists from using gait assessment methods in clinical practice was a lack of resources, including instruments, space, time, and funds.