title: Adenovirus vector-mediated RNA interference for the inhibition of human parvovirus B19 replication.
abstract: Human parvovirus B19 (B19V) has been considered to cause acute and chronic myocarditis, which is accompanied by endothelial dysfunction. Currently, no causative treatment option for B19V-infections is available. Since RNA interference (RNAi) has proven to be a highly potent antiviral approach, the aim of the current study was to develop an RNAi-based strategy to inhibit B19V replication. Three B19V-VP2-specific short hairpin RNAs (shRNAs) were designed and tested for their silencing activity in reporter assays and the expression cassette of the best one was introduced into an adenoviral shuttle vector (Ad5). B19V-permissive UT7/Epo-S1 cells were infected with B19V and the RNAi triggers were delivered by the adenoviral vector (Ad5shVP2) 24h thereafter. The shRNA targeting the B19V-VP2 gene significantly suppressed VP2 mRNA levels as determined by quantitative RT-PCR. Additionally, also the expression levels of the non-targeted non-structural B19V-NS1 mRNA were strongly reduced. Our results demonstrate that vector-mediated delivery of shRNA expression cassettes targeting the structural B19-VP2 gene is a suitable approach to inhibit B19V replication.
authors.affiliation: Department of Molecular Pathology, Institute of Pathology and Neuropathology, University Hospital of Tuebingen, 72076 Tuebingen, Germany.
authors.forename: Marius R G
authors.initials: MR
authors.lastname: Brandt
authors.forename: Ariane G
authors.initials: AG
authors.lastname: Kirste
authors.forename: Tanja
authors.initials: T
authors.lastname: Pozzuto
authors.forename: Steffen
authors.initials: S
authors.lastname: Schubert
authors.forename: Reinhard
authors.initials: R
authors.lastname: Kandolf
authors.forename: Henry
authors.initials: H
authors.lastname: Fechner
authors.forename: C-Thomas
authors.initials: CT
authors.lastname: Bock
authors.forename: Jens
authors.initials: J
authors.lastname: Kurreck
mesh_terms: D000260:Adenoviruses, Human / Q000235:genetics*; D000998:Antiviral Agents / Q000378:metabolism; D002460:Cell Line; D020869:Gene Expression Profiling; D005822:Genetic Vectors; D006801:Humans; D016732:Parvovirus B19, Human / Q000235:genetics / Q000502:physiology*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D014779:Virus Replication
chemical_list: D000998:Antiviral Agents; D034741:RNA, Small Interfering
keywords: Antiviral therapy; Parvovirus B19; RNA interference; Small interfering RNA