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Efficient transduction of myeloid cells by an HIV-1-derived lentiviral vector that packages the Vpx accessory protein

Gene Therapy volume 20, pages 514–520 (2013)Cite this article

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Abstract

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.

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Acknowledgements

We thank Antonia Follenzi for the pPGK.GFP plasmid and Kayleigh Taylor for technical assistance. The studies were funded by the National Institutes of Health (A1067059 and 5T32 A1007180). NRL is an Elizabeth Glazer Scientist of the Pediatric AIDS Foundation.

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  1. S Bobadilla and N Sunseri: These authors contributed equally to this work.

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  1. Department of Microbiology, New York University School of Medicine, New York, NY, USA

    S Bobadilla, N Sunseri & N R Landau

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  1. S Bobadilla
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Correspondence to N R Landau.

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Bobadilla, S., Sunseri, N. & Landau, N. Efficient transduction of myeloid cells by an HIV-1-derived lentiviral vector that packages the Vpx accessory protein. Gene Ther 20, 514–520 (2013). https://doi.org/10.1038/gt.2012.61

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