The effects of flaxseed, sardines, pearl millet, and holy basil (Oscimum sanctum) leaf meal on Single Comb White Leghorn (Forsgate strain) layer hen production traits, fatty acid composition of egg yolks, and egg quality characteristics were studied. Layer hens were allocated to 1 of 6 dietary treatments from 27 to 32 wk of age: control (treatment 1), control + 1 g of basil leaf meal (BLM) (treatment 2), control + 2 g of BLM (treatment 3), designer egg layer mash (DELM; treatment 4), DELM + 1 g of BLM (treatment 5), DELM + 2 g of BLM (treatment 6). No significant differences were found in egg production between dietary treatments, indicating that neither the designer diet nor the BLM had an adverse effect on egg production. The high energy level in the designer diet, because of the flaxseed oil and sardines, significantly reduced feed intake compared with the control diet. The egg weight was significantly higher in birds fed the designer diet compared with the egg weight in the control diet. The flaxseed oil and sardine oil, which are rich in n-3 polyunsaturated fatty acids, contributed to significantly increased n-3 polyunsaturated fatty acid levels in the egg yolks, with a proportionate reduction in the level of palmitic acid. Other fatty acids showed only minor variations. Flaxseed oil increased the level of α-linolenic acid, whereas fish oil increased the eicosapentaenoic acid and docosahexaenoic acid levels. The BLM at low levels of inclusion (1 and 2 g/kg) also contributed significantly to elevating the yolk n-3 polyunsaturated fatty acid levels, especially C18:3. Holy basil leaf meal is a good feed supplement for increasing the yolk α-linolenic acid level by reducing the yolk saturated fatty acid levels proportionately.