Streptozotocin (STZ) has been widely used to create diabetic retinopathy (DR) model in a variety of animals. The clinical features and pathological changes of the animal models of DR induced by STZ are similar to those of DR of human type 1 diabetes mellitus, so the animal models are frequently used to study the pathogenesis of DR and evaluate new pre-clinical anti-DR drugs. Mice are one of the best models to study DR because of small size, low cost, ease of operation and high success rate of induction. However, researchers also face many challenges in inducing mouse model of DR, such as reproducibility of the model and animal lethality by STZ. In the process of inducing animal DR model by STZ, we need to pay attention to several key factors, including the preparation of STZ, suitable dosage, route of administration, feeding method, and age, body mass and gender of the animals. This review analyzes the mechanism of STZ-induced DR, emphasizes the important processes of STZ-induced mouse DR model, and proposes the methods to reduce the lethality of STZ.