title: Efficient inhibition of porcine reproductive and respiratory syndrome virus replication by artificial microRNAs targeting the untranslated regions.
abstract: A robust artificial microRNA (amiRNA) strategy against porcine reproductive and respiratory syndrome virus (PRRSV) was developed by targeting the untranslated regions (UTRs). Six candidate amiRNAs targeting the 5' or 3' UTR were used for vector construction, and four effective amiRNAs were selected for further study using a vector transfection/virus infection assay. In cell cultures stably transfected with the four amiRNA vectors, expression of the sequence-specific amiRNAs was confirmed using poly(A)-tailed RT-PCR. After infection with three different PRRSV strains, the viral RNA genome and/or transcript were inhibited by ~90 % (semi-quantitative RT-PCR), and the viral titers were decreased by more than six log CCID(50) (viral titration assay) before day 3 postinfection. The potent anti-PRRSV effects lasted for at least 5 days. Sequence analysis showed that the amiRNA antiviral activities were not compromised by the presence of one or two mismatches in their binding targets. This work constitutes a step towards developing a more effective RNAi strategy against PRRSV.
authors.affiliation: College of Veterinary Medicine, Yangzhou University, 48 Wenhui Road, Yangzhou 225009, Jiangsu, People's Republic of China.
authors.forename: Bing
authors.initials: B
authors.lastname: Xia
authors.forename: Hongqin
authors.initials: H
authors.lastname: Song
authors.forename: Yang
authors.initials: Y
authors.lastname: Chen
authors.forename: Xinyu
authors.initials: X
authors.lastname: Zhang
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Xia
authors.forename: Huaichang
authors.initials: H
authors.lastname: Sun
mesh_terms: D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D015536:Down-Regulation; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D019318:Porcine Reproductive and Respiratory Syndrome / Q000821:virology*; D019316:Porcine respiratory and reproductive syndrome virus / Q000235:genetics* / Q000502:physiology; D034622:RNA Interference; D012367:RNA, Viral / Q000235:genetics* / Q000378:metabolism; D013552:Swine; D020506:Untranslated Regions; D014779:Virus Replication
chemical_list: D035683:MicroRNAs; D012367:RNA, Viral; D020506:Untranslated Regions