title: Efficient generation of sFat-1 transgenic rabbits rich in n-3 polyunsaturated fatty acids by intracytoplasmic sperm injection.
abstract: N-3 polyunsaturated fatty acids (n-3 PUFAs) have their first double bond at the third carbon from the methyl end of the fatty-acid chain and had been proven to be beneficial to human health. However, mammals cannot produce n-3 PUFAs by themselves because they lack the n-3 fatty-acid desaturase (Fat-1) gene. Thus, the possibility of producing sFat-1 transgenic rabbits was explored in this study. The transgenic cassette of pPGK1-sFat-1-CMV-EGFP was constructed and transgenic rabbit embryos were produced by intracytoplasmic sperm injection (ICSI). When 123 EGFP-positive embryos at the 2-8-cell stage were transplanted into the oviduct of four oestrous-synchronised recipients, two of them became pregnant and gave birth to seven pups. However, transfer of embryos into the uterus of oestrous-synchronised recipients and oviduct or uterus of oocyte donor rabbits did not result in pregnancy. The integration of the sFat-1 gene was confirmed in six of the seven live pups by PCR and Southern blot. The expression of the sFat-1 gene in the six transgenic pups was also detected by reverse transcription polymerase chain reaction (RT-PCR). Gas chromatography-mass spectrometry analysis revealed that transgenic rabbits exhibited an ~15-fold decrease in the ratio of n-6:n-3 PUFAs in muscle compared with wild-type rabbits and non-transgenic rabbits. These results demonstrate that sFat-1 transgenic rabbits can be produced by ICSI and display a low ratio of n-6:n-3 PUFAs.
authors.affiliation: State Key Laboratory for Conservation and Utilisation of Subtropical Agro-Bioresources, Guangxi University, Nanning 530004, China.
authors.forename: Shun
authors.initials: S
authors.lastname: Zhang
authors.affiliation: State Key Laboratory for Conservation and Utilisation of Subtropical Agro-Bioresources, Guangxi University, Nanning 530004, China.
authors.forename: Fenghua
authors.initials: F
authors.lastname: Lu
authors.affiliation: State Key Laboratory for Conservation and Utilisation of Subtropical Agro-Bioresources, Guangxi University, Nanning 530004, China.
authors.forename: Qingyou
authors.initials: Q
authors.lastname: Liu
authors.affiliation: State Key Laboratory for Conservation and Utilisation of Subtropical Agro-Bioresources, Guangxi University, Nanning 530004, China.
authors.forename: Yubing
authors.initials: Y
authors.lastname: Liu
authors.affiliation: State Key Laboratory for Conservation and Utilisation of Subtropical Agro-Bioresources, Guangxi University, Nanning 530004, China.
authors.forename: Xiaomei
authors.initials: X
authors.lastname: Guan
authors.affiliation: State Key Laboratory for Conservation and Utilisation of Subtropical Agro-Bioresources, Guangxi University, Nanning 530004, China.
authors.forename: Yingming
authors.initials: Y
authors.lastname: Wei
authors.affiliation: State Key Laboratory for Conservation and Utilisation of Subtropical Agro-Bioresources, Guangxi University, Nanning 530004, China.
authors.forename: Shijian
authors.initials: S
authors.lastname: Tan
authors.affiliation: State Key Laboratory for Conservation and Utilisation of Subtropical Agro-Bioresources, Guangxi University, Nanning 530004, China.
authors.forename: Deshun
authors.initials: D
authors.lastname: Shi
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001755:Blastocyst / Q000201:enzymology*; D046149:Embryo Culture Techniques / Q000662:veterinary; D004624:Embryo Transfer / Q000662:veterinary; D004790:Enzyme Induction; D044943:Fatty Acid Desaturases / Q000096:biosynthesis* / Q000235:genetics; D015525:Fatty Acids, Omega-3 / Q000378:metabolism*; D043371:Fatty Acids, Omega-6 / Q000378:metabolism; D005260:Female; D008401:Gas Chromatography-Mass Spectrometry / Q000662:veterinary; D005838:Genotype; D008297:Male; D008460:Meat; D018482:Muscle, Skeletal / Q000378:metabolism*; D010641:Phenotype; D011247:Pregnancy; D018873:Pregnancy Rate; D011817:Rabbits; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D020554:Sperm Injections, Intracytoplasmic / Q000662:veterinary*
chemical_list: D015525:Fatty Acids, Omega-3; D043371:Fatty Acids, Omega-6; D044943:Fatty Acid Desaturases; C078565:omega-3 fatty acid desaturase