Incorporation of dietary n-3 polyunsaturated fatty acids (PUFA) into bone may optimize bone development. The study objective was to use the fat-1 mouse, a transgenic model that synthesizes n-3 PUFA from n-6 PUFA, to determine if bone mineral density (BMD) and biomechanical bone strength were favourably modulated by lowering the n-6/n-3 PUFA ratio in vertebrae. Male and female wild-type and fat-1 mice were fed an AIN93-G diet containing 10% safflower oil from weaning through 12 weeks of age. Vertebrae BMD was determined by dual energy x-ray absorptiometry and peak load, a surrogate measure of fracture risk, was measured by a materials testing system. Vertebrae fatty acid composition was measured by gas liquid chromatography. At 12 weeks of age, vertebrae peak load was higher in fat-1 mice compared to wild-type (P = 0.026). Fat-1 mice also had lower n-6/n-3 PUFA ratio in vertebrae than wild-type (P < 0.001) and this ratio was negatively correlated with BMD and peak load (P = 0.005). Moreover, n-3 PUFA including alpha-linolenic acid, eicosapentaenoic acid and docosahexaenoic acid were positively correlated (P < 0.05) with BMD and peak load. Therefore, a lower vertebrae n-6/n-3 PUFA ratio is associated with stronger vertebrae and suggests a positive role for n-3 PUFA in bone development.