Eggs are a nutritious food, offering a balanced source of essential amino and fatty acids, minerals, and vitamins. Genetic and diet factors can modify hen egg traits. Thus, the effects of breed and feed on egg quality using two laying hens, Mos (autochthonous breed) and Isa Brown (commercial hybrid), and three feeds, commercial fodder (CF), corn/pea/triticale (CPT) and corn/wheat (CW), were investigated. Freshness parameters (egg weight, eggshell weight and thickness, albumen height, Haugh units and yolk color), chemical composition, color and textural parameters, as well as fatty acid profile, were assessed on a total of 288 eggs, from the two breeds. The samples were divided in 96 eggs, corresponding to each of the three dietary treatments. There were significant differences (p < 0.001) in albumen height and Haugh units, obtaining the highest values for Isa Brown genotype; meanwhile, laying hens fed with CF had the highest weight, as well as the greatest eggshell thickness. Cooked yolks of Isa Brown eggs presented the highest values of luminosity, while the yellowness was higher for Mos eggs. Regarding the texture of eggs, genotype was again the parameter having the greatest impact, obtaining higher values in hardness, gumminess and chewiness in those eggs from the Mos breed. Concerning egg chemical composition, it was affected by breed and diet type, but Mos eggs were characterized by a significantly (p < 0.001) higher contents of fat (9.53% vs. 7.58%), protein (12.31% vs. 11.66%) and ash (1.10% vs. 1.04%) than Isa Brown ones. Finally, diet type influenced the fatty acid profile, mainly affecting oleic and linoleic acids, which showed significantly (p < 0.05) highest values (42.90 and 14.66 g/100 g of total fatty acids) in CW and CF diets, respectively. Overall, breed and bird diet factors had a strong effect on egg quality and nutritional profile. Moreover, eggs from Mos hens had more attractive nutritional indices, and they could even be improved more by changing the diet.