Efficient transduction of myeloid cells by an HIV-1-derived lentiviral vector that packages the Vpx accessory protein.
Lentiviral vectors are widely used for the stable expression of genes and small hairpin RNA (shRNA)-mediated knockdown and are currently under development for clinical use in gene therapy. Pseudotyping of the vectors with VSV-G allows them to infect a wide range of cell types. However, myeloid cells, such as dendritic cells and macrophages, are relatively refractory to lentiviral vector transduction as a result of the myeloid-specific restriction factor, SAMHD1. SIVmac/HIV-2 and related viruses relieve the SAMHD1-mediated restriction by encoding Vpx, a virion-packaged accessory protein that induces the degradation of SAMHD1 upon infection. HIV-1 does not encode Vpx and cannot package the protein. We report the development of an HIV-1-based lentiviral vector in which the Vpx packaging motif has been placed in the p6 region of the Gag/Pol expression vector that is used to generate the lentiviral vector virions. The virions package Vpx in high copy number and infect myeloid cells with a two-log increase in titer. Transduction of dendritic cells with an shRNA against transportin-3 resulted in >90% knockdown of the encoding mRNA. The system can be applied to any HIV-based lentiviral vector and is useful for laboratory and clinical applications where the efficient transduction of myeloid cells is required.
Department of Microbiology, New York University School of Medicine, New York, NY, USA.
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Bobadilla
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Sunseri
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Landau
D003713:Dendritic Cells / Q000166:cytology; D015316:Genetic Therapy; D005822:Genetic Vectors; D015497:HIV-1 / Q000235:genetics; D015498:HIV-2 / Q000235:genetics; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D008264:Macrophages; D022423:Myeloid Cells / Q000166:cytology / Q000378:metabolism; D034741:RNA, Small Interfering; D014161:Transduction, Genetic; D054334:Viral Regulatory and Accessory Proteins / Q000235:genetics*
D034741:RNA, Small Interfering; C074286:VPX protein, Human immunodeficiency virus 2; D054334:Viral Regulatory and Accessory Proteins