title: Transient gene expression mediated by integrase-defective retroviral vectors.
abstract: Nonintegrating retroviral vectors were produced from a Moloney murine leukemia virus (MoMLV)-based retroviral vector system by introducing a point mutation into the integrase (IN) gene of the packaging plasmid. The efficacy of IN-defective retroviral vectors was measured through the transient expression of ZsGreen or luciferase in human cell lines. The IN-defective retroviral vectors could transduce target cells efficiently, but their gene expression was transient and lower than that seen with the integrating vectors. IN-defective retroviral vector gene expression decreased to background levels in fewer than 10 days. Southern blot analysis of transduced K562 cells confirmed the loss of a detectable vector sequence by 15 days. The residual integration activity of the IN-defective vector was 1000- to 10,000-fold lower than that of the integrating vector. These results demonstrate that the IN-defective retroviral vectors can provide a useful tool for efficient transient gene expression targeting of primary hematopoietic stem cells and lymphoid cells.
authors.affiliation: Center for Cell and Gene Therapy, Takara Bio, Seta3-4-1, Otsu, Shiga 520-2193, Japan.
authors.forename: Seung Shin
authors.initials: SS
authors.lastname: Yu
authors.forename: Kazuyuki
authors.initials: K
authors.lastname: Dan
authors.forename: Hideto
authors.initials: H
authors.lastname: Chono
authors.forename: Emi
authors.initials: E
authors.lastname: Chatani
authors.forename: Junichi
authors.initials: J
authors.lastname: Mineno
authors.forename: Ikunoshin
authors.initials: I
authors.lastname: Kato
mesh_terms: D018952:Antigens, CD34; D004279:DNA, Viral / Q000502:physiology; D015870:Gene Expression; D005822:Genetic Vectors; D006412:Hematopoietic Stem Cells; D006801:Humans; D019426:Integrases / Q000172:deficiency*; D020014:K562 Cells; D008979:Moloney murine leukemia virus / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D014161:Transduction, Genetic
chemical_list: D018952:Antigens, CD34; D004279:DNA, Viral; D019426:Integrases