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Improvement of lentiviral vector-mediated gene transduction by genetic engineering of the structural protein Pr55Gag

Gene Therapy volume 17, pages 1124–1133 (2010)Cite this article

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Abstract

The lentiviral vector is a promising tool for human gene therapy because of its ability to transduce genes into many cell types. However, one of the technical problems associated with the lentiviral vector is that lentiviral titers in current production systems are relatively low compared with the other viral vectors. In this study, we provide genetic evidence that the attachment of heterologous myristoylation (myr) signals on the amino-terminus of human immunodeficiency virus type 1 Pr55Gag (Gag) can increase the viral yield up to 10-fold, leading to the enhancement of gene transduction in many cell lines. The myr signal Gag constructs behaved similarly to the wild-type Gag in targeting to detergent-resistant membrane compartments, Vps4-dependence for viral budding, and virion morphology. However, the myr signal Gag constructs showed improved oligomerization efficiency as measured by bioluminescence resonance energy transfer in living cells, contributing to increased viral production and efficient activation of the viral protease responsible for virion maturation. The genetically modified Gag represents the next generation lentiviral vector, and should contribute to the success of many lentiviral vector applications.

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Acknowledgements

This work was supported by the Japan Health Science Foundation, the Japanese Ministry of Health, Labor and Welfare (H18-AIDS-W-003 to JK) and the Japanese Ministry of Education, Culture, Sports, Science and Technology (18689014 and 18659136 to JK).

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Authors and Affiliations

  1. AIDS Research Center, National Institute of Infectious Diseases, Tokyo, Japan

    T Aoki, S Shimizu, E Urano, Y Futahashi, M Hamatake, T Murakami, N Yamamoto & J Komano

  2. Department of Medicinal Chemistry, Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, Tokyo, Japan

    T Aoki & H Tamamura

  3. Graduate School of Infection Control Sciences, Kitasato University, Tokyo, Japan

    E Urano

  4. Department of Comprehensive Pathology, Aging and Developmental Sciences, Tokyo Medical and Dental University, Graduate School, Tokyo, Japan

    K Terashima

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  1. T Aoki
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  2. S Shimizu
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  3. E Urano
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  4. Y Futahashi
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  8. T Murakami
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  9. N Yamamoto
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  10. J Komano
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Correspondence to J Komano.

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Aoki, T., Shimizu, S., Urano, E. et al. Improvement of lentiviral vector-mediated gene transduction by genetic engineering of the structural protein Pr55Gag. Gene Ther 17, 1124–1133 (2010). https://doi.org/10.1038/gt.2010.61

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  • DOI: https://doi.org/10.1038/gt.2010.61

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