In spaceflight operations, flight controllers manage technical aspects of spaceflight control and spacecraft systems. The flight control team is a group subjected to shift work. Acute and chronic exposure to shift work has been associated with circadian misalignment, sleep impairment, and a negative impact on cognitive performance. This study aims to review the effects of shift work on sleep, circadian rhythms, mood, and cognitive performance of flight controllers during real and simulated spaceflight operations. Shift work during low-Earth orbit spaceflight missions is associated with a reduction in alertness, motivation, processing speed, and working memory performance efficiency. Flight controllers also report excessive insomnia and insufficient total sleep time. The development of shift work sleep disorder may be present in up to 40% of workers, especially among night and evening shift workers. Mars operations and Mars-simulated missions are associated with an impairment of visual-motor performance, working memory efficiency, and reaction time. Between 50%–87% of controllers can synchronize their circadian rhythm to a Mars day (24.65 hours), although this adaptation is not reflected in improved neurobehavioral performance.