title: High-titre retroviral vector system for efficient gene delivery into human and mouse cells of haematopoietic and lymphocytic lineages.
abstract: Genetically modified cells of haematopoietic and lymphocytic lineages could provide potentially curative treatments for a wide range of inherited and acquired diseases. However, this application is limited in mouse models by the low efficiency of lentiviral vectors. To facilitate the rapid production of high-titre helper-free retroviral vectors for enhanced gene delivery, multiple modifications to a prototype moloney murine leukemia virus (MoMLV)-derived vector system were made including adaptation of the vector system to simian virus 40 ori/T antigen-mediated episomal replication in packaging cells, replacement of the MoMLV 5' U3 promoter with a series of stronger composite promoters and addition of an extra polyadenylation signal downstream of the 3' long terminal repeat. These modifications enhanced vector production by 2-3 logs. High-titre vector stocks were tested for their ability to infect a variety of cells derived from humans and mice, including primary monocyte-derived macrophage cultures. Whilst the lentiviral vector was significantly restricted at the integration level, the MoMLV-based vector showed effective gene transduction of mouse cells. This high-titre retroviral vector system represents a useful tool for efficient gene delivery into human and mouse haematopoietic and lymphocytic cells, with particular application in mice as a small animal model for novel gene therapy tests.
authors.affiliation: Departments of Public Health Sciences and Microbiology, University of Hawaii at Manoa, Honolulu, HI 96822, USA.
authors.forename: Chengxiang
authors.initials: C
authors.lastname: Wu
authors.affiliation: Departments of Public Health Sciences and Microbiology, University of Hawaii at Manoa, Honolulu, HI 96822, USA.
authors.forename: Yuanan
authors.initials: Y
authors.lastname: Lu
mesh_terms: D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D005822:Genetic Vectors; D006801:Humans; D008214:Lymphocytes / Q000821:virology*; D008264:Macrophages / Q000821:virology*; D051379:Mice; D008979:Moloney murine leukemia virus / Q000235:genetics*; D011401:Promoter Regions, Genetic; D018741:Replication Origin; D013539:Simian virus 40 / Q000235:genetics; D014161:Transduction, Genetic; D019065:Virus Assembly; D014779:Virus Replication
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