The Micro-12 flight experiment was launched on SpaceX-15 and completed during berthing on the International Space Station. The goal of this experiment was to understand the effects of spaceflight and microgravity on the physiology of the model exoelectrogen Shewanella oneidensis MR-1. BioServe Fluid Processing Apparatus (FPA) and Group Activation Pack (GAP) hardware systems were used for both flight and ground control tests. Under spaceflight conditions, extracellular electron transfer (EET) rates were found to be significantly increased on insoluble substrates, while biofilm development appeared to be unchanged under the conditions tested; these processes are critical for microbial-assisted bioelectrochemical systems. Additionally, RNAseq analysis, proteomic profiling, and competitive mutant fitness profiling were performed to gain further understanding of microbial physiology under EET-respiring conditions during spaceflight. Overall, the results of the Micro-12 project support the idea that Shewanella oneidensis MR-1, in particular, and exoelectrogens in general could be useful chassis organisms for synthetic biology applications using microbial bioelectrochemical systems. These findings will assist bioengineering and synthetic biology development efforts harnessing the unique capabilities of exoelectrogens for life support and in situ resource utilization.