Construction of multiple shRNA vectors targeting PEDV and TGEV and production of transgenic SCNT porcine embryos in vitro

Jianwen CHEN; Kaiyuan PAN; Zhen CHEN; Biao DING; Dandan SONG; Wenbin BAO; Yunhai ZHANG

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Construction of multiple shRNA vectors targeting PEDV and TGEV and production of transgenic SCNT porcine embryos in vitro

机译：靶向PEDV和TGEV的多个shRNA载体的构建及体外转基因SCNT猪胚胎的产生

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Porcine viral diarrhea is an acute and highly contagious enteric disease of pigs that causes huge economic losses worldwide.Porcine epidemic diarrhea virus (PEDV) and transmissible gastroenteritis virus (TGEV) are the main pathogens responsible for piglet viral diarrhea.However,currently there is no specific drug available for the effective treatment of viral diarrhea.Therefore,it is necessary to seek an effective method to diminish PEDV and TGEV infection rates.RNA interference has been applied successfully to inhibit the virus replication.It provides a potential strategy for breeding resistant pigs.In this study,four promoters and four short hairpin RNA (shRNA) vectors with LoxP sites at each end of the selectable marker genes were constructed to target PEDV and TGEV.These vectors were then transfected into porcine fetal fibroblasts,G418 resistant transfectants were confirmed by PCR and transgenic SCNT porcine blastocysts were obtained.These results have paved the way for future production of marker-free transgenic resistant to PEDV and TEGV pigs by SCNT.

机译：猪病毒性腹泻是一种急性和高度传染性的猪肠道疾病，在全球范围内造成巨大的经济损失。猪流行性腹泻病毒（PEDV）和可传播的肠胃炎病毒（TGEV）是造成仔猪病毒性腹泻的主要病原体。因此，有必要寻求一种有效的方法来降低PEDV和TGEV的感染率。RNA干扰已成功应用于抑制病毒复制，为抗性猪的繁殖提供了潜在的策略。本研究构建了四个启动子和四个在选择标记基因两端具有LoxP位点的短发夹RNA（shRNA）载体，以靶向PEDV和TGEV，然后将这些载体转染到猪胎儿成纤维细胞中，确定了G418抗性转染子通过PCR，获得了转基因SCNT的猪囊胚，为今后的生产铺平了道路。 SCNT诱导无标记的转基因抗PEDV和TEGV猪。

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《农业科学与工程前沿（英文版）》 |2019年第1期|66-72|共7页
作者
Jianwen CHEN; Kaiyuan PAN; Zhen CHEN; Biao DING; Dandan SONG; Wenbin BAO; Yunhai ZHANG;
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Anhui Provincial Laboratory of Local Livestock and Poultry, Genetical Resource Conservation and Breeding, College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China;

Anhui Provincial Laboratory of Local Livestock and Poultry, Genetical Resource Conservation and Breeding, College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China;

Anhui Provincial Laboratory of Local Livestock and Poultry, Genetical Resource Conservation and Breeding, College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China;

Anhui Provincial Laboratory of Local Livestock and Poultry, Genetical Resource Conservation and Breeding, College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China;

Anhui Provincial Laboratory of Local Livestock and Poultry, Genetical Resource Conservation and Breeding, College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China;

College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China;

Anhui Provincial Laboratory of Local Livestock and Poultry, Genetical Resource Conservation and Breeding, College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China;

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Construction of multiple shRNA vectors targeting PEDV and TGEV and production of transgenic SCNT porcine embryos in vitro

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