A modified n-3 fatty acid desaturase gene from Caenorhabditis briggsae produced high proportion of DHA and DPA in transgenic mice.
The functions of polyunsaturated fatty acids (PUFAs) have been widely investigated. In mammals, levels of n-3 PUFAs are relatively low compared to those of n-6 PUFAs. Either a lack of n-3 PUFAs or an excess of n-6 PUFAs could potentially cause health problems in humans. Hence, methods to increase the amount of n-3 PUFAs in diet have been intensely sought. In this study, we demonstrated that the n-3 fatty acid desaturase gene (sFat-1) synthesized from revised and optimized codons based on roundworm Caenorhabditis briggsae genomic gene for enhanced expression in mammals was successfully expressed in Chinese hamster ovary (CHO) cells and significantly elevated cellular n-3 PUFA contents. We generated sFat-1 transgenic mice by introducing mammal expression vector DNAs containing the sFat-1 gene into regular mice through the method of microinjection. Fatty acid compositions were then altered and the levels of docosahexaenoic acid (DHA, 22:6n-3) and docosapentaenoic acid (DPA, 22:5n-3) were greatly increased in these transgenic mice. Various types of tissues in the transgenic mice produced many types of n-3 PUFAs, such as alpha-linolenic acid (ALA; 18:3n-3), eicosapentaenoic acid (EPA, 20:5n-3), DPA, and DHA, for example, muscle tissues of the transgenic mice contained 12.2% DHA, 2.0% DPA, and 23.1% total n-3 PUFAs. These research results demonstrated that the synthesized sFat-1 gene with modified and optimized codons from C. briggsae possess functional activity and greater capability of producing n-3 PUFAs, especially DHA and DPA, in transgenic mice.
Zhu
Guiming
G
Beijing Institute of Biotechnology, Beijing, China.
Chen
Hongxing
H
Wu
Xiaojie
X
Zhou
Yanrong
Y
Lu
Jianshen
J
Chen
Hong
H
Deng
Jixian
J
D000818:Animals; D016466:CHO Cells; D002107:Caenorhabditis / Q000201:enzymology / Q000235:genetics*; D029742:Caenorhabditis elegans Proteins / Q000502:physiology*; D006224:Cricetinae; D003412:Cricetulus; D004281:Docosahexaenoic Acids / Q000378:metabolism*; D044943:Fatty Acid Desaturases / Q000502:physiology*; D005231:Fatty Acids, Unsaturated / Q000378:metabolism*; D005260:Female; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D011401:Promoter Regions, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014018:Tissue Distribution
D029742:Caenorhabditis elegans Proteins; D005231:Fatty Acids, Unsaturated; C485184:enhanced green fluorescent protein; C466741:fat-1 protein, C elegans; D049452:Green Fluorescent Proteins; D004281:Docosahexaenoic Acids; D044943:Fatty Acid Desaturases; C026219:docosapentaenoic acid