Objective The basic biological, echocardiography and gene sequencing parameters of Slit2 gene overexpressing mice (Slit2-Tg mice) were collected and evaluated, which provided a reference for the application of Slit2-Tg mice in biomedical research. Methods Slit2-Tg and C57BL/6J mice were inbred. The genotypes of the mice were determined by a PCR method. The blood samples were collected for blood routine and blood biochemical tests. The tissues of main organs were collected for protein expression and pathological analysis. Echocardiography and transcriptome sequencing was carried for analyzing the heart function and gene expression, respectively. Results Litter size was significantly higher in Slit2-Tg mice than in C57BL/6J mice. Human Slit2 gene and protein expression were detected in the main organs of Slit2-Tg mice. Organ coefficient of spleen was increased significantly in Slit2-Tg mice, but the tissue structure appeared normal. There were significant changes in the counts of erythrocytes, platelets, eosinophils, and biochemistry of glucose, globulin, urea nitrogen, triglycerides, HDL, and atherosclerosis index. Echocardiography showed no significant differences in the morphology and function of the Slit2-Tg hearts except in the left ventricular anterior wall thickness at the end-diastolic state. Compared with C57BL/6J, 535 genes out of 17513 genes in the Slit2-Tg hearts were increased or decreased, mainly involving 15 biological process or signal transduction pathways. Conclusion This study has collected the biological parameters of Slit2-Tg mice and suggests that this model animal is suitable for the study of cardiovascular diseases.