This study was aimed at investigating the effect of dietary fat modulation via oil blend supplement (linseed oil 1.5/3.0%, sunflower oil 1.5/3.0%, 20 g FineX™NV1810) on the milk production, quality, fatty acid profile and gene (PPAR-α, SCD, ACAC, HMG-CoA synthase) expression of lactating Barbari goats. Eighteen goats were equally divided into three groups, where group A was given a basal diet, while group B and group C were offered two levels of oil blend in addition to the basal diet for 90 days. Oil blend supplementation significantly (P < 0.05) affected milk production, pH, titratable acidity, ash, milk solid-not-fat, and total soluble solids. Further, it significantly (P < 0.05) increased C4:0 C16:1, C18:0, C18:1n–9, C18:2n–6, C18:3n–3, MUFA, PUFA, and P/S ratio, while reduced the fatty acids C8:0 to C16:0, n–6/n–3 ratio, and SFA of milk fat. There was a positive effect (P < 0.05) of the oil blend supplement on the health and metabolic indices of milk fat. The oil supplementation levels and trial period affected the expression of the studied genes.