Background: The Democratic Republic of the Congo (DRC) witnessed the world’s second largest Ebola Virus Disease (EVD) outbreak killing 2290 individuals, with women disportionately infected (57% of all cases) despite no evidence on differential biological risk for EVD. We assessed if men and women differ in personal protective behaviors (vaccine acceptance, health-seeking behaviors, physical distancing) and the mediating role of gender differences in levels of EVD information and knowledge, perceived disease risk, and social relations. Methods: We conducted a multi-stage cluster survey of 1395 individuals in North Kivu between 20 April – 10 May, 2019, a period of high transmission in the sampled region of DRC. We conducted path analyses using a structural equation modeling framework to examine associations among variables as hypothesized. Findings: We found associations between gender and all EVD preventive behavioral outcomes, with evidence for the mediating role of EVD knowledge and belief in rumors. Men reported greater EVD accuracy compared to women. Greater EVD knowledge accuracy was associated with increases in vaccine acceptance, formal care seeking, and self-protection behaviors. Lower belief in rumors led to greater vaccine acceptance, and greater EVD information awareness led to increased adoption of self-protection behaviors. Interpretation: In any response to infectious disease, such as the current COVID-19 pandemic, it is critical to not only design gender-sensitive communication and vaccination strategies, but also to engage women and the community as a whole in the response.Funding: The Enhanced Learning and Research for Humanitarian Assistance (ELRHA) and United Nations Development Programme (UNDP) provided funding for the study.Declaration of Interest: We declare no competing interests.Ethical Approval: The survey was approved by Brigham and Women’s Hospital Human Research Committee and the Research Center on Democracy and Development in Africa, Free university of the Great Lakes Countries in the Democratic Republic of the Congo.