High-titre retroviral vector system for efficient gene delivery into human and mouse cells of haematopoietic and lymphocytic lineages.
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.
Wu
Chengxiang
C
Departments of Public Health Sciences and Microbiology, University of Hawaii at Manoa, Honolulu, HI 96822, USA.
Lu
Yuanan
Y
Departments of Public Health Sciences and Microbiology, University of Hawaii at Manoa, Honolulu, HI 96822, USA.
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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