Journal
JOURNAL OF GENERAL VIROLOGY
Volume 91, Issue -, Pages 1909-1918Publisher
MICROBIOLOGY SOC
DOI: 10.1099/vir.0.020255-0
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Funding
- Hawaii Community Foundation [20070438]
- National Institute of Health [S11 NS43499-01A1, MH079717-01A2]
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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.
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