We examined the genotype-phenotype interactions of Cyp51^+/2 mice carrying one functional allele of lanosterol 14ademethylase from cholesterol biosynthesis. No distinct developmental or morphological abnormalities were observed by routine visual inspection of Cyp51^+/2 and Cyp51+/+ mice and fertility was similar. We further collected a large data-set from female and male Cyp51^+/2 mice and controls fed for 16 weeks with three diets and applied linear regression modeling. We used 3 predictor variables (genotype, sex, diet), and 39 response variables corresponding to the organ characteristics (7), plasma parameters (7), and hepatic gene expression (25). We observed significant differences between Cyp51^+/2 and wildtype mice in organ characteristics and blood lipid profile. Hepatomegaly was observed in Cyp51^+/2 males, together with elevated total and low-density lipoprotein cholesterol. Cyp51^+/2 females fed high-fat, high-cholesterol diet were leaner and had elevated plasma corticosterone compared to controls. We observed elevated hepatocyte apoptosis, mitosis and lipid infiltration in heterozygous knockouts of both sexes. The Cyp51^+/2 females had a modified lipid storage homeostasis protecting them from weight-gain when fed high-fat high-cholesterol diet. Malfunction of one Cyp51 allele therefore initiates disease pathways towards cholesterol-linked liver pathologies and sex-dependent response to dietary challenge.