Diabetic retinopathy(DR)is one of the most common retinal complications of diabetes could cause irreversible loss of central vision in the working-age population. Current studies showed that systemic risk factors, inflammatory response, and oxidative stress played a central role in the development of DR. Although traditional sequencing methods have provided valuable insights into the pathogenesis of DR, offering crucial guidance for clinical diagnosis and treatment, they still possess certain limitations. In recent years, the emerging single-cell RNA sequencing technology(scRNA-seq)has enabled precise analysis of mRNA transcriptomes at the single-cell level. This technique accurately identifies novel cell subtypes in retinal diseases, detects rare cells, and reveals intercellular heterogeneity. It contributes to elucidating the pathogenesis and development of retinal diseases, and facilitates exploration of gene regulatory relationships associated with these disorders to provide valuable insights for future precision medicine. This article reviews the technology of single-cell sequencing and its application in DR research. It also explores the mechanisms of different types of cells associated with DR, aiming to enhance the utilization of scRNA-seq in DR research and identify potential therapeutic targets to improve clinical diagnosis and treatment of DR.