title: Development of sheep kidney cells with increased resistance to different subgenotypes of BVDV-1 by RNA interference.
abstract: Bovine viral diarrhea virus (BVDV) should be a ubiquitous viral pathogen to the cattle and sheep industry. This pathogen is responsible for severe economic losses. We previously showed that plasmid-mediated dual short hairpin RNA (shRNA) efficiently inhibit BVDV replication in bovine kidney epithelial (MDBK) cells. In this study, we delivered the dual shRNA system to sheep fibroblasts and generated transgenic cell colonies. These transgenic fibroblasts were further used for somatic cell nuclear transfer (SCNT). Three lambs were born at full term, but perished soon after birth. Integration of shRNA into the genome of cloned sheep was confirmed by PCR and expression of shRNA in transgenic sheep was confirmed by real-time PCR. Kidney epithelial cells were isolated from transgenic sheep and challenged with multiple BVDV subgenotypes (BVDV-1a, BVDV-1b and BVDV-1c). The dual shRNA expressed in transgenic kidney epithelial cells significantly inhibited BVDV replication in a cross-resistance manner. Our results showed that transgenic RNAi might be a useful tool for preparation of transgenic animals with increased resistance to BVDV.
authors: College of Life Sciences, Shihezi University, Shihezi, Xinjiang 832003, China.
authors: Wei
authors: W
authors: Ni
authors: College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang 832003, China.
authors: Jun
authors: J
authors: Qiao
authors: College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang 832003, China.
authors: Qiman
authors: Q
authors: Ma
authors: College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang 832003, China.
authors: Jiangde
authors: J
authors: Wang
authors: College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang 832003, China.
authors: Dawei
authors: D
authors: Wang
authors: College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang 832003, China.
authors: Xinxia
authors: X
authors: Zhao
authors: College of Life Sciences, Shihezi University, Shihezi, Xinjiang 832003, China.
authors: Yang
authors: Y
authors: Cao
authors: College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang 832003, China.
authors: Qifeng
authors: Q
authors: Li
authors: College of Life Sciences, Shihezi University, Shihezi, Xinjiang 832003, China. Electronic address: shengwei.hu@aggiemail.usu.edu.
authors: Shengwei
authors: S
authors: Hu
authors: College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang 832003, China.
authors: Chuangfu
authors: C
authors: Chen
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D002460:Cell Line; D030241:Diarrhea Virus 1, Bovine Viral / Q000235:genetics*; D060467:Disease Resistance / Q000235:genetics; D004847:Epithelial Cells / Q000821:virology; D053652:Nuclear Transfer Techniques; D018182:Pestivirus Infections / Q000473:pathology / Q000517:prevention & control* / Q000662:veterinary*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000627:therapeutic use; D060888:Real-Time Polymerase Chain Reaction; D012756:Sheep / Q000821:virology; D017361:Viral Nonstructural Proteins / Q000235:genetics*; D014764:Viral Proteins / Q000235:genetics*
chemical_list: C507353:Npro protein, bovine viral diarrhea virus; D034741:RNA, Small Interfering; D017361:Viral Nonstructural Proteins; D014764:Viral Proteins
keywords: BVDV; RNAi; SCNT; Transgenic sheep