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Animal tissue composition is determinant in the quality of animal products and is influenced by several factors including diet, genetic type, age and sex. Diet effect on Iberian pig's tissue composition is a relevant issue with productive and economic implications, not well understood to date. In this work we evaluated the effect of the diet fatty acid composition of fattening Iberian pigs, with respect to different tissues FA composition and transcription of three key genes coding for lipid metabolism enzymes (SCD, ME1 and FASN). Forty Torbiscal male pigs were subjected to three different diets: saturated fat (S), monounsaturated fat (M) and polyunsaturated fat (P) during the fattening period. Adipose tissue and liver fatty acid composition show great differences for P animals respect to S and M groups, which show a similar fatty acid profile. Regarding the diet effect on gene expression several effects are detected in the different tissues, suggesting a higher expression of lipogenic enzyme genes on the M group, and lower in the P group. These gene expression results do not explain the higher ability of S group for the endogenous synthesis of monounsaturated FA, which can be deduced from the tissue composition analyses.