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ABSTRACT A 90‐day feeding study in rats was conducted to evaluate the subchronic oral toxicity of genetically modified (GM) DAS‐81419–2 soybean. Wistar rats were fed with diets containing toasted soybean meal produced from DAS‐81419–2 soybean grain that expresses the Cry1F, Cry1Ac, and Pat proteins or containing conventional soybean at doses of 30.0%, 15.0%, 7.5%, or 0% (control group) for 90 consecutive days. The general behavior, body weight and food consumption were observed. At the middle and end of the experiment, blood, serum, and urine samples were collected for biochemical assays. At the conclusion of the study, the internal organs were weighed and histopathological examination was completed. The rats exhibited free movement and shiny coats without any abnormal symptoms or abnormal secretions in their noses, eyes, or mouths. There were no adverse effects on body weight in GM soybean groups and conventional soybean groups. No biological differences in hematological, biochemical, or urine indices were observed. No significant differences in relative organ weights were detected between the experimental groups and the control group. No histopathological changes were observed. Under the conditions of this study, DAS‐81419–2 soybean did not cause any treatment‐related effects in Wistar rats following 90 days of dietary administration. HIGHLIGHTSEvaluate the subchronic oral toxicity of genetically modified DAS‐81419–2 soybean.In the study, the rats' behavior, body weight, food consumption were observed.The blood, serum, and urine samples were collected for biochemical assays.The internal organs histopathological examination was completed.The GM DAS‐81419–2 soybean is substantially equivalent with non‐GM soybean.