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This study examined the changes in student learning outcomes as a result of participation in a large enrollment introductory biology course that implemented a novel educational model: Learning Assistants (LAs). LA-led activities presented new material and engaged students in discussions of personal relevance replacing passive, lecture-based lessons. Our results demonstrate that students in the LA program outperformed their counterparts on exams. This effect was moderated by both first-time-in-college status (FTIC students in LA sections outperformed their counterparts) and gender (female students in LA sections outperformed their counterparts). Both female and FTIC students performed significantly better when their interactions with LAs occurred in an active learning space designed for collaboration, rather than a traditional lecture hall.