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Protein transduction by pseudotyped lentivirus-like nanoparticles

Gene Therapy volume 18, pages 936–941 (2011)Cite this article

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Abstract

A simple, efficient and reproducible method to transduce proteins into mammalian cells has not been established. Here we describe a novel protein transduction method based on a lentiviral vector. We have developed a method to package several thousand foreign protein molecules into a lentivirus-like nanoparticle (LENA) and deliver them into mammalian cells. In this proof-of-concept study, we used β-lactamase (BlaM) as a reporter molecule. The amino-terminus of BlaM was fused to the myristoylation signal of lyn, which was placed upstream of the amino-terminus of Gag (BlaM-gag-pol). By co-transfection of plasmids encoding BlaM-gag-pol and vesicular stomatitis virus-G (VSV-G) into 293T cells, LENA were produced containing BlaM enzyme molecules as many as Gag per capsid, which has been reported to be ∼5000 molecules, but lacking the viral genome. Infection of 293T and MT-4 cells by VSV-G-pseudotyped BlaM-containing LENA led to successful transduction of BlaM molecules into the cell cytoplasm, as detected by cleavage of the fluorescent BlaM substrate CCF2-AM. LENA-mediated transient protein transduction does not damage cellular DNA, and the preparation of highly purified protein is not necessary. This technology is potentially useful in various basic and clinical 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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  1. T Aoki and K Miyauchi: These authors contributed equally to this work.

Authors and Affiliations

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

    T Aoki, K Miyauchi, E Urano, R Ichikawa & J Komano

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  1. T Aoki
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  2. K Miyauchi
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  3. E Urano
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  4. R Ichikawa
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  5. J Komano
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Correspondence to J Komano.

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Aoki, T., Miyauchi, K., Urano, E. et al. Protein transduction by pseudotyped lentivirus-like nanoparticles. Gene Ther 18, 936–941 (2011). https://doi.org/10.1038/gt.2011.38

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