Aims: The aim of this study was to evaluate the effectiveness of E-mail-based intervention on safe crossing behaviors among Isfahan University of Medical Science students, in year 2014 in Iran. Materials and Methods: In over all, 78 boy-students with 22.33 ± 1.65 years were participated in an E-mail-based intervening study. They divided into intervention group (n = 38) and control group (n = 40) by use of random allocation. The instrument of data collection included questionnaire of theory of planned behavior constructs and traffic behaviors. Questionnaires were filled out by both groups at the baseline and 6 months after the educational intervention. The safe crossing educational intervention was conducted within 1-month through E-mail service for the intervention group. The control group received no education. Results: Results showed no significant differences between groups in mean of attitude toward safe crossing, subjective norms (SNs), perceived behavioral control (PBC), intention and safe crossing behaviors at baseline (P > 0.05). MANCOVA test demonstrated that there is a significant difference between groups after the educational intervention with adjusting the effect of age (P = 0.001, F = 31.144, Wilks lambda = 0.313). According to the ANCOVA test, after educational intervention, mean of positive attitude toward safe crossing, PBC, intention and behaviors related to safe crossing were increased in intervention group in comparing control group (P < 0.005). Conclusion: E-mail-based intervention can promote safe crossing behaviors among college students. Conducting further studies with focusing on SNs related to safe crossing is suggested.