Abstract
Current techniques for making transgenic mice are cumbersome, requiring trained personnel, costly infrastructure and collection of many zygotes from mice that are then killed. We developed a reproducible nonterminal technique for transfecting genes in undifferentiated spermatogonia through in vivo electroporation of the testis; about 94% of male mice electroporated with different transgenes successfully sired transgenic pups. Such electroporated males provide a valuable resource for continuous production of transgenic founders for more than a year.
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Acknowledgements
We thank S. Babajko (Institut National de la Santé et de la Recherche Médicale, Paris), S. Chattopadhyay (National Center for Cell Science, Pune, India), Y. Junming (University of Tennessee, Memphis) and J. DeJong (University of Texas at Dallas) for providing constructs. We are thankful for technical inputs of A. Usmani and M. Gautam at various steps. We thank S.K. Basu, A. Surolia, K.V.S. Rao and T.M. Plant for their helpful suggestions. We acknowledge the help of V. Bal, A. Mukhopadhyay, K. Sarda, M. Shrivastava, P. Seth, A. Basu and R. Das for immunohistochemistry and stem cell culture–related work. This work was funded by the National Institute of Immunology (India) and Department of Biotechnology. We are also thankful to Indo-US Contraceptive and Reproductive Health Research program.
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Dhup, S., Majumdar, S. Transgenesis via permanent integration of genes in repopulating spermatogonial cells in vivo. Nat Methods 5, 601–603 (2008). https://doi.org/10.1038/nmeth.1225
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DOI: https://doi.org/10.1038/nmeth.1225
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