Transgenesis of humanized fat1 promotes n-3 polyunsaturated fatty acid synthesis and expression of genes involved in lipid metabolism in goat cells

The n - 3 fatty acid desaturase gene fatl codes for the n 3 desaturase enzyme, which can convert n 6 polyunsaturated fatty acids (PUFAs) to n 3 PUFAs. The n 3 PUFAs are essential components required for normal cellular function and have preventive and therapeutic effects on many diseases. Goat is an important domestic animal for human consumption of meat and milk. To elevate the concentrations of n 3 PUFAs and examine the regulatory mechanism of fatl in PUFA metabolism in goat cells, we successfully constructed a humanized fatl expression vector and confirmed the efficient expression of fatl in goat ear skin -derived fibroblast cells (GEFCs) by qRT-PCR and Western blot analysis. Fatty acid analysis showed that fatl overexpression significantly increased the levels of total n 3 PUFAs and decreased the levels of total n 6 PUFAs in GEFCs. In addition, qRTPCR results indicate that the FADS1 and FADS2 desaturase genes, ELOV2 and ELOV5 elongase genes, ACO and CPT1 oxidation genes, and PPARa and PPARy transcription factors are up-regulated, and transcription factors of SREBPI c gene are down-regulated in the fatl transgenic goat cells. Overall, fatl-overexpression resulted in an increase in the n 3 fatty acids and altered expression of PUFA synthesis related genes in GEFCs. This work lays a foundation for both the production of fatl transgenic goats and further study of the mechanism of fatl function in the PUFAs metabolism. (C) 2015 Elsevier B.V. All rights reserved.