title: Inhibition of porcine reproductive and respiratory syndrome virus replication with exosome-transferred artificial microRNA targeting the 3' untranslated region.
abstract: Porcine reproductive and respiratory syndrome (PRRS) is an economically important swine disease. As part of the development of RNA interference (RNAi) strategy against the disease, in this study a recombinant adenovirus (rAd) expressing the artificial microRNA (amiRNA) targeting the 3' untranslated region (UTR) was used to investigate the exosome-mediated amiRNA transfer from different pig cell types to porcine alveolar macrophages (PAMs). Quantitative RT-PCR showed that the sequence-specific amiRNA was expressed in and secreted via exosomes from the rAd-transduced pig kidney cell line PK-15, PAM cell line 3D4/163, kidney fibroblast cells (PFCs) and endometrial endothelial cells (PEECs) with different secretion efficiencies. Fluorescent microscopy revealed that the dye-labeled amiRNA-containing exosomes of different cell origins were efficiently taken up by all of the five types of pig cells tested, including primary PAMs. Quantitative RT-PCR showed that the amiRNA-containing exosomes of different cell origins were taken up by primary PAMs in both time- and dose-dependent manners. Both quantitative RT-PCR and viral titration assays showed that the exosome-delivered amiRNA had potent anti-viral effects against three different PRRSV strains. These data suggest that the exosomes derived from pig cells could serve as an efficient miRNA transfer vehicle, and that the exosome-delivered amiRNA had potent anti-viral effects against different PRRSV strains.
authors.affiliation: College of Veterinary Medicine, Jiangsu Co-Innovation Center for Prevention and Control of Important Animal infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, China.
authors.forename: Li
authors.initials: L
authors.lastname: Zhu
authors.affiliation: College of Veterinary Medicine, Jiangsu Co-Innovation Center for Prevention and Control of Important Animal infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, China.
authors.forename: Liping
authors.initials: L
authors.lastname: Bao
authors.affiliation: College of Veterinary Medicine, Jiangsu Co-Innovation Center for Prevention and Control of Important Animal infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, China.
authors.forename: Xinyu
authors.initials: X
authors.lastname: Zhang
authors.affiliation: College of Veterinary Medicine, Jiangsu Co-Innovation Center for Prevention and Control of Important Animal infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, China.
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Xia
authors.affiliation: College of Veterinary Medicine, Jiangsu Co-Innovation Center for Prevention and Control of Important Animal infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, China. Electronic address: sunh@yzu.edu.cn.
authors.forename: Huaichang
authors.initials: H
authors.lastname: Sun
mesh_terms: D020413:3' Untranslated Regions; D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D000998:Antiviral Agents / Q000378:metabolism*; D002478:Cells, Cultured; D004705:Endocytosis; D055354:Exosomes / Q000378:metabolism*; D015870:Gene Expression; D005822:Genetic Vectors; D035683:MicroRNAs / Q000378:metabolism*; D019316:Porcine respiratory and reproductive syndrome virus / Q000235:genetics* / Q000502:physiology*; D060888:Real-Time Polymerase Chain Reaction; D013552:Swine; D019562:Viral Load; D014779:Virus Replication / Q000187:drug effects*
chemical_list: D020413:3' Untranslated Regions; D000998:Antiviral Agents; D035683:MicroRNAs
keywords: Anti-viral effect; Artificial microRNA; Exosome-mediated transfer; Porcine reproductive and respiratory syndrome virus