Chicken eggs are considered one of the most common high nutritive value animal protein foods sources consumed all over the world. Recently, with the increasing of health-conscious there is a growing interest in producing and consuming functional foods especially for individuals suffering from chronic diseases. The current study aimed to investigate the effect of dietary linseed oil (LO) inclusion, as enriched-source of omega-3 polyunsaturated fatty acids, on egg production performance and egg yolk cholesterol content, blood hematology and biochemical composition of commercial laying hens. A total of 180 Hy-Line brown laying hens 36-week-old were randomly divided into four groups (9 replicates x 5 birds each). The four groups were fed a basal diet formulated to contain; 0, 2, 4 or 6% LO for six weeks. Results showed a significant improvement in egg production performance with dietary LO inclusion ranged from 3 to 4%. Data revealed a significant reduction in egg yolk cholesterol content by 15, 17 and 21% for the 2, 4 and 6% LO groups, respectively, compared to the 0% LO group. Meanwhile, blood high-density lipoprotein was significantly increased, while the low-and very-low-density lipoprotein cholesterols were significantly decreased by LO treatment, compared to the 0% LO group. Dietary LO did not show any adverse effects on the measured blood hematological or plasma biochemical parameters of the layers. It can be concluded that inclusion of LO into layer's diets can be safely used to enhance egg production. Furthermore, LO inclusion might have a beneficial impact on reducing egg cholesterol content which can be considered as an adding value for consumer's general health. The economic efficiency evaluation reviled that diet fortified by 6% LO is the most efficient diet in respect of the current Saudi Arabia market's prices.