title: Efficient transient genetic manipulation in vitro and in vivo by prototype foamy virus-mediated nonviral RNA transfer.
abstract: Vector systems based on different retroviruses are widely used to achieve stable integration and expression of transgenes. More recently, transient genetic manipulation systems were developed that are based on integration- or reverse transcription-deficient retroviruses. Lack of viral genome integration is desirable not only for reducing tumorigenic potential but also for applications requiring transient transgene expression such as reprogramming or genome editing. However, all existing transient retroviral vector systems rely on virus-encoded encapsidation sequences for the transfer of heterologous genetic material. We discovered that the transient transgene expression observed in target cells transduced by reverse transcriptase-deficient foamy virus (FV) vectors is the consequence of subgenomic RNA encapsidation into FV particles. Based on this initial observation, we describe here the establishment of FV vectors that enable the efficient transient expression of various transgenes by packaging, transfer, and de novo translation of nonviral RNAs both in vitro and in vivo. Transient transgene expression levels were comparable to integrase-deficient vectors but, unlike the latter, declined to background levels within a few days. Our results show that this new FV vector system provides a useful, novel tool for efficient transient genetic manipulation of target tissues by transfer of nonviral RNAs.
authors.affiliation: Institut für Virologie, Medizinische Fakultät "Carl Gustav Carus", Technische Universität Dresden, Dresden, Germany; CRTD/DFG-Center for Regenerative Therapies Dresden, Cluster of Excellence, Technische Universität Dresden, Dresden, Germany.
authors.forename: Martin V
authors.initials: MV
authors.lastname: Hamann
authors.affiliation: Institut für Virologie, Medizinische Fakultät "Carl Gustav Carus", Technische Universität Dresden, Dresden, Germany; CRTD/DFG-Center for Regenerative Therapies Dresden, Cluster of Excellence, Technische Universität Dresden, Dresden, Germany.
authors.forename: Nicole
authors.initials: N
authors.lastname: Stanke
authors.affiliation: Institut für Virologie, Medizinische Fakultät "Carl Gustav Carus", Technische Universität Dresden, Dresden, Germany; CRTD/DFG-Center for Regenerative Therapies Dresden, Cluster of Excellence, Technische Universität Dresden, Dresden, Germany.
authors.forename: Erik
authors.initials: E
authors.lastname: Müllers
authors.affiliation: Institut für Virologie, Medizinische Fakultät "Carl Gustav Carus", Technische Universität Dresden, Dresden, Germany; CRTD/DFG-Center for Regenerative Therapies Dresden, Cluster of Excellence, Technische Universität Dresden, Dresden, Germany.
authors.forename: Kristin
authors.initials: K
authors.lastname: Stirnnagel
authors.affiliation: Institut für Virologie, Medizinische Fakultät "Carl Gustav Carus", Technische Universität Dresden, Dresden, Germany; CRTD/DFG-Center for Regenerative Therapies Dresden, Cluster of Excellence, Technische Universität Dresden, Dresden, Germany.
authors.forename: Sylvia
authors.initials: S
authors.lastname: Hütter
authors.affiliation: CRTD/DFG-Center for Regenerative Therapies Dresden, Cluster of Excellence, Technische Universität Dresden, Dresden, Germany.
authors.forename: Benedetta
authors.initials: B
authors.lastname: Artegiani
authors.affiliation: CRTD/DFG-Center for Regenerative Therapies Dresden, Cluster of Excellence, Technische Universität Dresden, Dresden, Germany.
authors.forename: Sara
authors.initials: S
authors.lastname: Bragado Alonso
authors.affiliation: CRTD/DFG-Center for Regenerative Therapies Dresden, Cluster of Excellence, Technische Universität Dresden, Dresden, Germany.
authors.forename: Federico
authors.initials: F
authors.lastname: Calegari
authors.affiliation: Institut für Virologie, Medizinische Fakultät "Carl Gustav Carus", Technische Universität Dresden, Dresden, Germany; CRTD/DFG-Center for Regenerative Therapies Dresden, Cluster of Excellence, Technische Universität Dresden, Dresden, Germany. Electronic address: dirk.lindemann@tu-dresden.de.
authors.forename: Dirk
authors.initials: D
authors.lastname: Lindemann
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D005347:Fibroblasts / Q000166:cytology / Q000821:virology*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D066298:In Vitro Techniques; D051379:Mice; D012313:RNA / Q000378:metabolism*; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism; D016092:Spumavirus / Q000235:genetics* / Q000378:metabolism; D014161:Transduction, Genetic; D019076:Transgenes
chemical_list: D012313:RNA; D012194:RNA-Directed DNA Polymerase