Abstract
Until today, gene transfer to germ cells has been attempted by a variety of methods including microinjection, embryonic stem cell-mediated transfection, virus mediated transfection, lipofection (1), microparticle bombardment (2), and sperm mediated transformation (3). In vivo gene electroporation (EP) now is a viable option for gene transfer purposes as demonstrated by strong shortterm gene expression and long-term expression after gene transfer (4,5 and unpublished). In vivo gene EP is simple and convenient. Adequate development and differentiation of transfected cells can be maintained in the in vivo environment. Furthermore, it can be applied, in principle, to any types of cells and tissues so long as the target is accessible. The advantages of in vivo EP over other nonviral methods such as lipofection and microparticle bombardment are: possible tissue damage is less; there is no limitation of DNA to be transfected at a time; and DNA can be transferable to cells deep inside the target tissue.
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References
Northlop, J. P., Ringlod, G. M., and Danielsen, M. (1987) Lipofection: A highly efficient, lipid-mediated DNA-transfection procedure. Proc. Natl. Acad. Sci. U.S.A. 84, 7413–7417.
Williams, R. S., Johnston, S. A., Reidy, M., de Vit, M. J., McElligott, S. G., and Sanford, J. C. (1991) Introduction of foreign genes into tissues of living mice by DNA-coated microprojectiles. Proc. Natl. Acad. Sci. U.S.A. 88, 2726–2730.
Lauria, A. and Gandolfi, F. (1993) Recent advances in sperm cell mediated gene transfer. Mol. Reprod. Dev. 36, 255–257.
Nishi, T., Yoshizato, K., Yamashiro, S., Takeshima, H., Sato, K., Hamada, K., Kitamura, I., Yoshimura, T., Saya, H., Kuratsu, J., and Ushio, Y. (1996) Highefficiency in vivo gene transfer using intraarterial plasmid DNA injection following in vivo electroporation. Cancer Res. 56, 1050–1055.
Muramatsu, T., Nakamura, A., and Park, H.-M. (1998) In vivo electroporation: A powerful and convenient means of nonviral gene transfer to tissues of living animals. Int. J. Mol. Med. 1, 55–62.
Muramatsu, T., Shibata, O., and Okumura, J. (1997) Effect of self-replication DNA sequences of Epstein-Barr virus on the expression of a foreign gene transfected in vivo to the mouse testis. Anim. Sci. Technol. Jpn. 68, 650–653.
Muramatsu, T., Shibata, O., and Okumura, J. (1997) Foreign gene expression in the mouse testis by localized in vivo gene transfer. Biochem. Biophys. Res. Commun. 233, 45–49.
Nakamura, A., Okumura, J., and Muramatsu, T. (1998) Quantitative analysis of transcriptional activity of viral and hybrid promoters in bovine preimplantation embryos. Mol. Reprod. Dev. 49, 368–373.
Muramatsu, T., Mizutani, Y., Ohmori, Y., and Okumura J. (1996) In vivo electroporation: a convenient method for gene transfer to testicular cells in mice. Anim. Sci. Technol. Jpn. 67, 975–982.
Okabe, M., Ikawa, M., Kominami, K., Nakanishi, T., and Nishimune, Y. (1997) ‘Green mice’ as a source of ubiquitous green cells. FEBS Lett. 407, 313–319.
Shoji-Tanaka, A., Mizuochi, T., and Komuro, K. (1994) Gene transfer using purified retroviral integrase. Biochem. Biophys. Res. Commun. 203, 1756–1764.
Hauer, C. A., Getty, R. R., and Tykocinski, M. L. (1989) Epstein-Barr virus episome-based promoter function in human myeloid cells. Nucleic Acids Res. 17, 1989–2003.
Pothier, F., Ouellet, M., Julien, J.-P., and Guerin, S. L. (1992) An improved CAT assay for promoter analysis in either transgenic mice or tissue culture cells. DNA Cell Biol. 11, 83–90.
Sanders, M. M. and McKnight, G. S. (1988) Positive and negative regulatory elements control the steroid-responsive ovalbumin promoter. Biochemistry 27, 6550–6557.
Hofmann, G. A. (1995) Instrumentation. Methods Mol. Biol. 47, 27–45.
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Muramatsu, T. (2000). In Vivo Gene Electroporation in the Mouse Testis. In: Jaroszeski, M.J., Heller, R., Gilbert, R. (eds) Electrochemotherapy, Electrogenetherapy, and Transdermal Drug Delivery. Methods in Molecular Medicine, vol 37. Humana Press. https://doi.org/10.1385/1-59259-080-2:349
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DOI: https://doi.org/10.1385/1-59259-080-2:349
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