RNAi-based inhibition of porcine reproductive and respiratory syndrome virus replication in transgenic pigs

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• 作者：Li Li ; Qiuyan Li ; Yonghua Bao ; Jinxiu Li ; Zhisheng Chen ; Xiuling Yu ; Yaofeng Zhao ; Kegong Tian ; Ning Li
• 关键词：Porcine reproductive and respiratory syndrome virus (PRRSV) ; RNA interference ; Somatic cell nuclear transplantation (SCNT) ; Transgenic pig
• 刊名：Journal of Biotechnology
• 出版年：10 February, 2014
• 年：2014
• 卷：171
• 期：Complete
• 页码：17-24
• 全文大小：1395 K

文摘

Porcine reproductive and respiratory syndrome (PRRS) is an economically devastating viral disease causing heavy losses to the swine industry worldwide. Many studies have shown that transient delivery of small interfering RNA (siRNA) or adenovirus-mediated RNA interfere (RNAi) could potentially inhibit porcine reproductive and respiratory syndrome virus (PRRSV) replication in vivo and in vitro. Here, we applied RNAi to produce transgenic (TG) pigs that constitutively expressed PRRSV-specific siRNA derived from small hairpin RNA (shRNA). First, we evaluated siRNA expression in the founding and F1 generation pigs and confirmed stable transmission. Then, we detected the expression of IFN-尾 and protein kinase R (PKR) and found no difference among TG, non-transgenic (NTG), and wild-type pigs. Lastly, the F1 generation pigs, including TG and NTG piglets, were challenged with 3 脳 10^4.5 TCID₅₀ of JXA1, a highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV). Our results showed that the in vivo siRNA expression substantially reduced the serum HP-PRRSV titers and increased survival time by 3 days when TG pigs were compared with the NTG controls. These data suggested that RNAi-based genetic modification might be used to breed viral-resistant livestock with stable siRNA expression with no complications of siRNA toxicity.