title: RNAi-based inhibition of porcine reproductive and respiratory syndrome virus replication in transgenic pigs.
abstract: 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-β 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 3×10⁴·⁵ 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.
authors: State Key Laboratories of Agrobiotechnology, College of Biological Science, China Agricultural University, Beijing 100193, China; Department of Molecular Biology, College of Biological Science, China Agricultural University, Beijing 100193, China.
authors: Li
authors: L
authors: Li
authors: State Key Laboratories of Agrobiotechnology, College of Biological Science, China Agricultural University, Beijing 100193, China; Department of Molecular Biology, College of Biological Science, China Agricultural University, Beijing 100193, China.
authors: Qiuyan
authors: Q
authors: Li
authors: State Key Laboratories of Agrobiotechnology, College of Biological Science, China Agricultural University, Beijing 100193, China; Department of Molecular Biology, College of Biological Science, China Agricultural University, Beijing 100193, China.
authors: Yonghua
authors: Y
authors: Bao
authors: State Key Laboratories of Agrobiotechnology, College of Biological Science, China Agricultural University, Beijing 100193, China; Department of Molecular Biology, College of Biological Science, China Agricultural University, Beijing 100193, China.
authors: Jinxiu
authors: J
authors: Li
authors: State Key Laboratories of Agrobiotechnology, College of Biological Science, China Agricultural University, Beijing 100193, China; Department of Molecular Biology, College of Biological Science, China Agricultural University, Beijing 100193, China.
authors: Zhisheng
authors: Z
authors: Chen
authors: OIE Porcine Reproductive and Respiratory Syndrome Reference Laboratory, China Animal Disease Control Center, No. 20 Maizidian Road, Chaoyang District, Beijing 100125, China.
authors: Xiuling
authors: X
authors: Yu
authors: State Key Laboratories of Agrobiotechnology, College of Biological Science, China Agricultural University, Beijing 100193, China; Department of Molecular Biology, College of Biological Science, China Agricultural University, Beijing 100193, China.
authors: Yaofeng
authors: Y
authors: Zhao
authors: OIE Porcine Reproductive and Respiratory Syndrome Reference Laboratory, China Animal Disease Control Center, No. 20 Maizidian Road, Chaoyang District, Beijing 100125, China; National Research Center for Veterinary Medicine, Cuiwei Road, High-Tech. District, Luoyang, Henan Provice, China.
authors: Kegong
authors: K
authors: Tian
authors: State Key Laboratories of Agrobiotechnology, College of Biological Science, China Agricultural University, Beijing 100193, China; Department of Molecular Biology, College of Biological Science, China Agricultural University, Beijing 100193, China. Electronic address: ninglcau@cau.edu.cn.
authors: Ning
authors: N
authors: Li
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics*; D002478:Cells, Cultured; D005260:Female; D016676:Macrophages, Alveolar; D008297:Male; D053652:Nuclear Transfer Techniques; D019318:Porcine Reproductive and Respiratory Syndrome / Q000235:genetics / Q000517:prevention & control; D019316:Porcine respiratory and reproductive syndrome virus / Q000235:genetics*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D016019:Survival Analysis; D013552:Swine / Q000235:genetics
chemical_list: D034741:RNA, Small Interfering
keywords: Porcine reproductive and respiratory syndrome virus (PRRSV); RNA interference; Somatic cell nuclear transplantation (SCNT); Transgenic pig