Exoskeletons can assist humans during squatting and the assistance has the potential to reduce the physical demands. Although several squat assistance methods are available, the effect of personalized assistance on physical effort has not been examined. We hypothesize that personalized assistance will reduce the physical effort of squatting. We developed a human-in-the-loop Bayesian optimization scheme to minimize the metabolic cost of squatting using a unilateral ankle exoskeleton. The optimization identified subject-specific assistance parameters for ascending and descending during squatting and took 15.8 min on average to converge. The subject-specific optimized condition reduced metabolic cost by 19.9% and rectus femoris muscle activity by 28.7% compared to the condition without the exoskeleton with a higher probability of improvement compared to a generic condition. In an additional study with two participants, the personalized condition presented higher metabolic cost reduction than the generic condition. These reductions illustrate the importance of personalized ankle assistance using an exoskeleton for squatting, a physically intensive activity, and suggest that such a method can be applied to minimize the physical effort of squatting. Future work can investigate the effect of personalized squat assistance on fatigue and the potential risk of injury.