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Highly efficient gene transfer into murine liver achieved by intravenous administration of naked Epstein–Barr virus (EBV)-based plasmid vectors

Gene Therapy volume 8, pages 1508–1513 (2001)Cite this article

Abstract

Naked plasmid DNA (pDNA) injection could become an alternative procedure to viral and nonviral gene delivery systems. We have previously shown that Epstein–Barr virus (EBV)-based plasmid vectors containing the EBV nuclear antigen 1 (EBNA1) gene and the oriP sequence enable quite high and long-lasting expression in various in vitro and in vivo transfection systems. The EBV-based plasmids were intravenously injected into mice via their tail vein under high pressure. A large amount of the marker gene product was expressed in the liver; as much as 320 μg of luciferase was demonstrated per gram of liver at 8 to 24 h after a single injection with 10 μg of DNA. More than 70% of liver cells stained with X-gal when β-gal gene was transferred. The expression level was significantly higher than that obtained by conventional pDNA lacking the EBNA1 gene and oriP. On day 35 after the transfection, the expression from the EBV-based plasmid was approximately 100-fold stronger than the conventional pDNA gene expression. Both the EBNA1 gene and oriP are a prerequisite for the augmentation of the transfection efficiency. These results suggest that the intravascular transfection with naked EBV-based plasmid may provide a quite efficient, simple and convenient means to transduce therapeutic genes in vivo into the liver. Gene Therapy (2001) 8, 1508–1513.

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Acknowledgements

We thank Dr Jun-Ichi Miyazaki (Department of Nutrition and Physiological Chemistry, Osaka University Medical School) for his generous gift of the CAG promoter and Dr Wilfred TV Germeraad (Crucell, Leiden, The Netherlands) for critical reading of the manuscript. This research was supported by a grant-in-aid for scientific research from the Ministry of Education, Science, Sports and Culture, Japan and a grant from the Japanese Research Foundation for Clinical Pharmacology.

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Authors and Affiliations

  1. Department of Microbiology, Research Institute for Neurological Disease and Geriatrics, Kyoto Prefectural University of Medicine, Kyoto, Japan

    FD Cui, T Kishida, H Asada, K Yasutomi, E Satoh, J Imanishi & O Mazda

  2. Department of Orthopedics, Research Institute for Neurological Disease and Geriatrics, Kyoto Prefectural University of Medicine, Kyoto, Japan

    S Ohashi & T Kubo

  3. Department of Pathology and Applied Neurobiology, Research Institute for Neurological Disease and Geriatrics, Kyoto Prefectural University of Medicine, Kyoto, Japan

    S Fushiki

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Cui, F., Kishida, T., Ohashi, S. et al. Highly efficient gene transfer into murine liver achieved by intravenous administration of naked Epstein–Barr virus (EBV)-based plasmid vectors. Gene Ther 8, 1508–1513 (2001). https://doi.org/10.1038/sj.gt.3301551

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