The Depression Anxiety Stress Scales-21 (DASS-21) has three 7-item subscales (depression, anxiety, and stress). The current study aims assess the gender-based measurement invariance of the DASS-21 questionnaire in a Chinese university student sample from five different cities. The sample was composed of 13208 participants (62.3% female, mean age of 19.7 years, and SD age = 1.8). Multi-group confirmatory factor analysis supported full measurement invariance for the three subscales. The findings support the measurement invariance of DASS-21 scores across gender. Future research on the DASS should include additional validation across ethnicities and testing of all versions of the DASS.