Biol Reprod Keystone Symposia Conference on Frontiers in Reproductive Biology & Regulation of Fertility.
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BOR - Papers in Press, published online ahead of print June 9, 2004.
Biol Reprod 2004, 10.1095/biolreprod.104.031294
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BIOLOGY OF REPRODUCTION 71, 1202–1207 (2004)
DOI: 10.1095/biolreprod.104.031294
© 2004 by the Society for the Study of Reproduction, Inc.

Reproductive Technology

Transgenic Mice Produced by Retroviral Transduction of Male Germ Line Stem Cells In Vivo1

Mito Kanatsu-Shinohara3, Shinya Toyokuni4, and Takashi Shinohara2,3

Horizontal Medical Research Organization3 Department of Pathology and Biology of Diseases,4 Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan

Spermatogonial stem cells are the only stem cells in the postnatal body that can transmit parental genetic information to the offspring, making them an attractive target cell population for animal transgenesis. Although transgenic mice and rats were recently produced by retrovirus transduction of these cells in vitro, with transplantation of the transduced cells into infertile recipients, the difficulty of restoring fertility and preparing recipients using spermatogonial transplantation limits practical application of the technique. In this article, we describe a novel approach for producing transgenic animals by transducing spermatogonial stem cells in vivo using a retrovirus vector. Microinjection of retrovirus into immature seminiferous tubules resulted in the direct transduction of spermatogonial stem cells in situ, and the animals produced transgenic offspring after mating with females. Transgenic mice were produced in C57BL/6, BALB/C, A, and C3H backgrounds, with an average efficiency of 2.8%. The transgene was transmitted stably and expressed in the next generation. The technique overcomes the drawback of the in vitro-transduction approach, and will be useful as a novel method for producing transgenic animals as well as providing a means for analyzing the self-renewal and differentiation processes of spermatogonial stem cells in vivo.

1 Support for this research was provided by the Ministry of Education, Culture, Sports, Science and Technology of Japan.

2 Correspondence: Takashi Shinohara, Horizontal Medical Research Organization, Graduate School of Medicine, Kyoto University Yoshida-Konoe, Sakyo-ku, Kyoto 606-8501, Japan. FAX: 81 75 753 9306; takashi{at}mfour.med.kyoto-u.ac.jp






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Copyright © 2004 by the Society for the Study of Reproduction.