title: Hybrid lentivirus-transposon vectors with a random integration profile in human cells.
abstract: Gene delivery by human immunodeficiency virus type 1 (HIV-1)-based lentiviral vectors (LVs) is efficient, but genomic integration of the viral DNA is strongly biased toward transcriptionally active loci resulting in an increased risk of insertional mutagenesis in gene therapy protocols. Nonviral Sleeping Beauty (SB) transposon vectors have a significantly safer insertion profile, but efficient delivery into relevant cell/tissue types is a limitation. In an attempt to combine the favorable features of the two vector systems we established a novel hybrid vector technology based on SB transposase-mediated insertion of lentiviral DNA circles generated during transduction of target cells with integrase (IN)-defective LVs (IDLVs). By construction of a lentivirus-transposon hybrid vector allowing transposition exclusively from circular viral DNA substrates, we demonstrate that SB transposase added in trans directs efficient transposon mobilization from DNA circles in vector-transduced cells. Both transfected plasmid DNA and transduced IDLVs can serve as the source of active transposase. Most important, we demonstrate that the SB transposase overrides the natural lentiviral integration pathway and directs vector integration less frequently toward transcriptional units, resulting in a random genomic integration profile. The novel hybrid vector system combines the attractive features of efficient gene delivery by viral transduction and a safer genomic integration profile by DNA transposition.
authors.affiliation: Department of Human Genetics, University of Aarhus, Denmark.
authors.forename: Nicklas H
authors.initials: NH
authors.lastname: Staunstrup
authors.forename: Brian
authors.initials: B
authors.lastname: Moldt
authors.forename: Lajos
authors.initials: L
authors.lastname: Mátés
authors.forename: Palle
authors.initials: P
authors.lastname: Villesen
authors.forename: Maria
authors.initials: M
authors.lastname: Jakobsen
authors.forename: Zoltán
authors.initials: Z
authors.lastname: Ivics
authors.forename: Zsuzsanna
authors.initials: Z
authors.lastname: Izsvák
authors.forename: Jacob Giehm
authors.initials: JG
authors.lastname: Mikkelsen
mesh_terms: D002460:Cell Line; D005822:Genetic Vectors / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D019426:Integrases / Q000235:genetics; D016086:Lentivirus / Q000235:genetics*; D014161:Transduction, Genetic / Q000379:methods*; D019895:Transposases / Q000235:genetics; D016662:Virus Integration / Q000235:genetics*
chemical_list: D019426:Integrases; D019895:Transposases; C494471:sleeping beauty transposase, human