The effects of different concentrations (30, 50, and 70 kPa) and time (1 or 3 hr) of short‐term CO2 gas treatment on the quality, safety, and shelf life of “Charlotte” strawberry were evaluated during storage at 4°C. Color attributes were better maintained in CO2 treated strawberries while firmness increased significantly and maintained throughout the storage in 70 kPa CO2 treatments compared to untreated control. Total soluble solid, pH, and titratable acidity did not show remarkable changes among the treatments. Total anthocyanins accumulation was significantly (p ≤ .05) lower in fruits exposed to 70 kPa CO2 for 1 hr than that of control but insignificantly lower with other CO2 treatments at the end of storage while total phenolic content was almost unaffected by CO2 treatment. Lower number of microbial population and better maintenance of sensory qualities were found in strawberries treated with 70 kPa CO2; therefore, marketability extended by about 4 and 2 days compared to control and other CO2 treatments, respectively. Practical applications These results suggest that the physicochemical, microbial, and sensory quality of strawberries were better maintained in fruits exposed to 70 kPa CO2 either for 1 or 3 hr before storage than sample stored directly. The results of this study could be useful for maintaining the quality and extending the shelf life of “Charlotte” strawberry during storage or export at low temperature condition.