The selection of the leading limb during gait initiation in individuals with unilateral transtibial amputations can significantly affect various biomechanical parameters. However, there is currently no established recommendation for the suitable leading limb in this population. The systematic review was aimed to propose a preferred leading limb for gait initiation in individuals with unilateral transtibial amputations based on biomechanical parameters. Databases including Google Scholar, PubMed, Science Direct, and ISI Web of Knowledge, were searched. The first selection criterion was based on abstracts and titles to address the research question. A total of seven studies were included in this review, and the Downs and Black's checklist was used by three researchers to assess the risk of bias. The review included a total of 61 adults with unilateral transtibial amputations, with a mean age range of 41 to 64.43 years. The confidence level of the included studies was poor, and the observational cohort was the most common study design (n = 5). Most of the studies were not replicable. Four of the included studies recommended the prosthetic limb as the preferred leading limb. Individuals with unilateral transtibial amputations may experience biomechanical benefits, including a more normal center of pressure path, reduced limb loading, and increased ankle energy generation, when leading with their prosthetic limb during gait initiation. However, further research is necessary to establish a more conclusive recommendation for the preferred leading limb in this population.