title: Transgenesis of humanized fat1 promotes n-3 polyunsaturated fatty acid synthesis and expression of genes involved in lipid metabolism in goat cells.
abstract: The n-3 fatty acid desaturase gene fat1 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 fat1 in PUFA metabolism in goat cells, we successfully constructed a humanized fat1 expression vector and confirmed the efficient expression of fat1 in goat ear skin-derived fibroblast cells (GEFCs) by qRT-PCR and Western blot analysis. Fatty acid analysis showed that fat1 overexpression significantly increased the levels of total n-3 PUFAs and decreased the levels of total n-6 PUFAs in GEFCs. In addition, qRT-PCR results indicate that the FADS1 and FADS2 desaturase genes, ELOV2 and ELOV5 elongase genes, ACO and CPT1 oxidation genes, and PPARa and PPARγ transcription factors are up-regulated, and transcription factors of SREBP-1c gene are down-regulated in the fat1 transgenic goat cells. Overall, fat1-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 fat1 transgenic goats and further study of the mechanism of fat1 function in the PUFAs metabolism.
authors.affiliation: Jiangsu Livestock Embryo Engineering Laboratory, Nanjing Agricultural University, Nanjing 210095, China. Electronic address: fanyixuan@njau.edu.cn.
authors.forename: Yixuan
authors.initials: Y
authors.lastname: Fan
authors.affiliation: Jiangsu Livestock Embryo Engineering Laboratory, Nanjing Agricultural University, Nanjing 210095, China. Electronic address: 853748688@qq.com.
authors.forename: Caifang
authors.initials: C
authors.lastname: Ren
authors.affiliation: Jiangsu Agri-animal Husbandry Vocational College, Taizhou 225300, China. Electronic address: 747551898@qq.com.
authors.forename: Zhibo
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Jiangsu Livestock Embryo Engineering Laboratory, Nanjing Agricultural University, Nanjing 210095, China. Electronic address: 772674111@qq.com.
authors.forename: Ruoxin
authors.initials: R
authors.lastname: Jia
authors.affiliation: Jiangsu Livestock Embryo Engineering Laboratory, Nanjing Agricultural University, Nanjing 210095, China. Electronic address: 563476324@qq.com.
authors.forename: Dan
authors.initials: D
authors.lastname: Wang
authors.affiliation: Jiangsu Livestock Embryo Engineering Laboratory, Nanjing Agricultural University, Nanjing 210095, China. Electronic address: zhangyanli@njau.edu.cn.
authors.forename: Yanli
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Jiangsu Livestock Embryo Engineering Laboratory, Nanjing Agricultural University, Nanjing 210095, China. Electronic address: 645081243@qq.com.
authors.forename: Guomin
authors.initials: G
authors.lastname: Zhang
authors.affiliation: Jiangsu Livestock Embryo Engineering Laboratory, Nanjing Agricultural University, Nanjing 210095, China. Electronic address: wanyongjie@njau.edu.cn.
authors.forename: Yongjie
authors.initials: Y
authors.lastname: Wan
authors.affiliation: Jiangsu Livestock Embryo Engineering Laboratory, Nanjing Agricultural University, Nanjing 210095, China. Electronic address: hmr@njau.edu.cn.
authors.forename: Mingrui
authors.initials: M
authors.lastname: Huang
authors.affiliation: Jiangsu Livestock Embryo Engineering Laboratory, Nanjing Agricultural University, Nanjing 210095, China. Electronic address: caeet@njau.edu.cn.
authors.forename: Feng
authors.initials: F
authors.lastname: Wang
mesh_terms: D000123:Acetyltransferases / Q000235:genetics; D000818:Animals; D030801:Animals, Genetically Modified; D015820:Cadherins / Q000235:genetics* / Q000378:metabolism; D002478:Cells, Cultured; D000090943:Delta-5 Fatty Acid Desaturase; D044943:Fatty Acid Desaturases / Q000235:genetics; D015525:Fatty Acids, Omega-3 / Q000096:biosynthesis*; D005347:Fibroblasts / Q000378:metabolism; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D006041:Goats / Q000235:genetics*; D050356:Lipid Metabolism / Q000235:genetics*; D047493:PPAR alpha / Q000235:genetics; D047495:PPAR gamma / Q000235:genetics; D051780:Sterol Regulatory Element Binding Protein 1 / Q000235:genetics
chemical_list: D015820:Cadherins; D000090943:Delta-5 Fatty Acid Desaturase; C502403:FAT1 protein, human; D015525:Fatty Acids, Omega-3; D047493:PPAR alpha; D047495:PPAR gamma; D051780:Sterol Regulatory Element Binding Protein 1; D044943:Fatty Acid Desaturases; C000716847:FADS1 protein, human; D000123:Acetyltransferases
keywords: Ear fibroblast cells; Goat; fat1; n−3 polyunsaturated fatty acids; n−6 polyunsaturated fatty acids