Objectives: The timing of protein intake over the day on muscle mass and strength gains have received interest in the literature. Thus, the aim of this systematic review is to analyze clinical studies that evaluated the acute effects of pre-sleep protein consumption on overnight muscle protein synthesis and the chronic effects on muscle mass and strength. Designs: Systematic review. Methods: A literature search was conducted up to June 2020 according to PRISMA statement and nine articles were included to analyze. Results: The consumption of 20-40 g of casein approximately 30 min before sleep stimulates whole-body protein synthesis rates over a subsequent overnight period in young and elderly men (preceded or not by resistance exercise, respectively). In addition, pre-sleep protein consumption can augment the muscle adaptive response (muscle fiber cross-sectional area, strength and muscle mass) during 10-12 weeks of resistance exercise in young, but not in elderly men. Conclusions: Based on current evidence, the consumption of 20-40 g of casein approximately 30 min before sleep improves protein synthetic response during an overnight recovery period in healthy young adult men, with possible positive effects on muscle mass and strength following prolonged resistance exercise. In elderly, despite the initial evidence regarding the pre-sleep protein enhances overnight muscle protein synthesis rates, the current available evidence is limited precluding to conclude about the chronic effects on skeletal muscle mass or strength. These conclusions need to be taken with caution due to uneven protein intakes between experimental groups. Therefore, more data are needed before further considering pre-sleep protein as an effective nutritional intervention.