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Objective To identify differences in biomechanics during gait in individuals with acute and persistent low back pain compared with back-healthy controls. Design Systematic review Data Sources A search was conducted in PubMed, CINAHL, SPORTDiscus, and PsycINFO in June 2019 and was repeated in December 2020. Eligibility criteria Studies were included if they reported biomechanical characteristics of individuals with and without low back pain during steady-state or perturbed walking and running. Biomechanical data included spatiotemporal, kinematic, kinetic, and electromyography variables. The reporting quality and potential for bias of each study was assessed. Data were pooled where possible to compare the standardized mean differences (SMD) between groups. Results Ninety-seven studies were included. Two studies investigated acute pain and the rest investigated persistent pain. Eight studies investigated running gait. 20% of studies had high reporting quality/low risk of bias. In comparison with back-healthy controls, individuals with persistent low back pain walked more slowly (SMD -0.59 [95% CI -0.77 to -0.42]) and with shorter stride length (-0.38 [-0.60 to -0.16]). There were no differences in the amplitude of motion in the thoracic or lumbar spine, pelvis, or hips in individuals with LBP. During walking, coordination of motion between the thorax and the lumbar spine/pelvis was significantly more in-phase in the LBP groups (-0.60 [-0.90 to -0-.30]), and individuals with LBP exhibited greater amplitude of activation in the paraspinal muscles (0.52 [0.23 to 0.80]). Summary/Conclusion There is moderate to strong evidence that individuals with persistent LBP demonstrate impairments in walking gait compared with back-healthy controls.