Sperm-mediated gene transfer
Marialuisa Lavitrano A B , Marco Busnelli A , Maria Grazia Cerrito A , Roberto Giovannoni A , Stefano Manzini A and Alessia Vargiolu AA Dipartimento di Scienze Chirurgiche e Terapia Intensiva, Università di Milano, Bicocca, 20052 Milano, Italy.
B Corresponding author. Email: marialuisa.lavitrano@unimib.it
Reproduction, Fertility and Development 18(2) 19-23 https://doi.org/10.1071/RD05124
Submitted: 21 September 2005 Accepted: 21 September 2005 Published: 14 December 2005
Abstract
Since 1989, a new method for the production of transgenic animals has been available, namely sperm-mediated gene transfer (SMGT), based on the intrinsic ability of sperm cells to bind and internalise exogenous DNA molecules and to transfer them into the oocyte at fertilisation. We first described the SMGT procedure in a small animal model, with high efficiency reported in the mouse. In addition, we successfully adapted and optimised the technique for use in large animals; it was, in fact, highly efficient in the generation of human decay accelerating factor transgenic pig lines, as well as multigene transgenic pigs in which three different reporter genes, namely enhanced green fluorescent protein, enhanced blue fluorescent protein and red fluorescent protein, were introduced. The major benefits of the SMGT technique were found to be its high efficiency, low cost and ease of use compared with other methods. Furthermore, SMGT does not require embryo handling or expensive equipment. Sperm-mediated gene transfer could also be used to generate multigene transgenic pigs that would be of benefit as large animal models for medical research, for agricultural and pharmaceutical applications and, in particular, for xenotransplantation, which requires extensive genetic manipulation of donor pigs to make them suitable for grafting to humans.
Extra keywords: sperm cells, transgenic animals.
Acknowledgments
The authors’ work reported herein was funded by the Italian Ministry of Agriculture (DM 581/7240/96, DM 564/7240/97, DM 404/7240/99, DM 36/7303/02) and by the Italian Minister of Research and University (DD 21.09.99, n462 ric). The authors thank Elena Giovannoni for assistance and the care of animals.
Brackett, B. G. , Baranska, W. , Sawichi, W. , and Koprowski, H. (1971). Uptake of heterologous genome by mammalian spermatozoa and its transfer to ova through fertilization. Proc. Natl Acad. Sci. USA 68, 353–357.
Lavitrano, M. , Forni, M. , Varzi, V. , Pucci, L. , and Bacci, M. L. , et al. (1997a). Sperm-mediated gene transfer: production of pigs transgenic for a human regulator of complement activation. Transplant. Proc. 29, 3508–3509.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
![](../../media/share/OpenURL_Image.gif)
Lavitrano, M. , Maione, B. , Forte, E. , Francolini, M. , Sperandio, S. , Testi, R. , and Spadafora, C. (1997b). The interaction of sperm cells with exogenous DNA: a role of CD4 and major histocompatibility complex class II molecules. Exp. Cell Res. 233, 56–62.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
![](../../media/share/OpenURL_Image.gif)
Lavitrano, M. , Bacci, M. L. , Forni, M. , Lazzereschi, D. , and Di Stefano, C. , et al. (2002). Production by sperm-mediated gene transfer of human decay accelerating factor (hDAF) transgenic pigs for xenotransplantation. Proc. Natl Acad. Sci. USA 99, 14 230–14 235.
| Crossref | GoogleScholarGoogle Scholar |
![](../../media/share/OpenURL_Image.gif)
Lavitrano, M. , Forni, M. , Bacci, M. L. , Di Stefano, C. , Varzi, V. , Wang, H. , and Seren, E. (2003). Sperm mediated gene transfer in pig: selection of donor boars and optimization of DNA uptake. Mol. Reprod. Dev. 64, 284–291.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
![](../../media/share/OpenURL_Image.gif)
Magnano, A. R. , Giordano, R. , Moscufo, N. , Baccetti, B. , and Spadafora, C. (1998). Sperm/DNA interaction: integration of foreign DNA sequences in the mouse sperm genome. J. Reprod. Immunol. 41, 187–196.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
![](../../media/share/OpenURL_Image.gif)
Maione, B. , Lavitrano, M. , Spadafora, C. , and Kiessling, A. A. (1998). Sperm-mediated gene transfer in mice. Mol. Reprod. Dev. 50, 406–409.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
![](../../media/share/OpenURL_Image.gif)
Mori, T. , Guo, M. W. , Mori, E. , Shindo, Y. , Mori, N. , Fukuda, I. , and Mori, T. (1990). Expression of class II major histocompatibility complex antigen on sperm and its role in fertilization. Am. J. Reprod. Immunol. 24, 9–10.
| PubMed |
![](../../media/share/OpenURL_Image.gif)
Muller, F. , Ivics, Z. , Erdelyi, F. , Papp, T. , Varadi, L. , Horvath, L. , and Maclean, N. (1992). Introducing foreign genes into fish eggs with electroporated sperm as a carrier. Mol. Mar. Biol. Biotechnol. 1, 276–281.
| PubMed |
![](../../media/share/OpenURL_Image.gif)
Niemann, H. , and Kues, W. A. (2003). Application of transgenesis in livestock for agriculture and biomedicine. Anim. Reprod. Sci. 79, 291–317.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
![](../../media/share/OpenURL_Image.gif)
Rieth, A. , Pothier, F. , and Sirard, M. A. (2000). Electroporation of bovine spermatozoa to carry DNA containing highly repetitive sequences into oocytes and detection of homologous recombination events. Mol. Reprod. Dev. 57, 338–345.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
![](../../media/share/OpenURL_Image.gif)
Rottmann, O. J. , Antes, R. , Hoefer, P. , and Maierhofer, G. (1991). Liposomes mediate gene transfer via spermatozoa into avian egg cells. J. Anim. Breed. Genet. 109, 64–70.
![](../../media/share/OpenURL_Image.gif)
Schellander, K. , Peli, J. , Schmall, F. , and Brem, G. (1995). Artificial insemination in cattle with DNA-treated sperm. Anim. Biotech. 6, 41–50.
![](../../media/share/OpenURL_Image.gif)
Sciamanna, I. , Piccoli, S. , Barberi, L. , Zaccagnini, G. , Magnano, A. R. , Giordano, R. , Campedelli, P. , Hodgson, C. , Lorenzini, R. , and Spadafora, C. (2000). DNA dose and sequence dependence in sperm-mediated gene transfer. Mol. Reprod. Dev. 56, 301–305.
| Crossref | GoogleScholarGoogle Scholar |
![](../../media/share/OpenURL_Image.gif)
Shemesh, M. , Gurevich, M. , Harel-Markowitz, E. , Benvenisti, L. , Shore, L. S. , and Stram, Y. (2000). Gene integration into bovine sperm genome and its expression in transgenic offspring. Mol. Reprod. Dev. 56(Suppl. 2), 306–308.
| Crossref | GoogleScholarGoogle Scholar |
![](../../media/share/OpenURL_Image.gif)
Smith, K. , and Spadafora, C. (2005). Sperm-mediated gene transfer: applications and implications. Bioessays 27, 551–562.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
![](../../media/share/OpenURL_Image.gif)
Spadafora, C. (1998). Sperm cells and foreign DNA: a controversial relation. BioEssays 20, 955–964.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
![](../../media/share/OpenURL_Image.gif)
Sperandio, S. , Lulli, V. , Bacci, M. L. , Forni, M. , Maione, B. , Spadafora, C. , and Lavitrano, M. (1996). Sperm mediated DNA transfer in bovine and swine species. Anim. Biotechnol. 7, 59–77.
![](../../media/share/OpenURL_Image.gif)
Tsai, H. J. , Lai, C. H. , and Yang, H. S. (1997). Sperm as a carrier to introduce an exogenous DNA fragment into the oocyte of Japanese abalone (Haliotis divorsicolor suportexta). Transgenic Res. 6, 85–95.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
![](../../media/share/OpenURL_Image.gif)
Umemoto, Y. , Sasaki, S. , Kojima, Y. , Kubota, H. , Kaneko, T. , Hayashi, Y. , and Kohri, K. (2005). Gene transfer to mouse testes by electroporation and its influence on spermatogenesis. J. Androl. 26, 264–271.
| PubMed |
![](../../media/share/OpenURL_Image.gif)
Wall, R. J. (1999). Sperm-mediated gene transfer: advances in sperm cell research and applications. Transgenic Res. 8, 313–315.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
![](../../media/share/OpenURL_Image.gif)
Wall, R. J. (2001). Pronuclear microinjection. Cloning Stem Cells 3, 209–220.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
![](../../media/share/OpenURL_Image.gif)
Wall, R. J. (2002). New gene transfer methods. Theriogenology 57, 189–201.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
![](../../media/share/OpenURL_Image.gif)
Webster, N. L. , Forni, M. , Bacci, M. L. , Giovannoni, R. , and Razzini, R. , et al. (2005). Multi-transgenic pigs expressing three fluorescent proteins produced with high efficiency by sperm mediated gene transfer. Mol. Reprod. Dev. 72, 68–76.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
![](../../media/share/OpenURL_Image.gif)
Wu, G. M. , Nose, K. , Mori, E. , and Mori, T. (1990). Binding of foreign DNA to mouse sperm mediated by its MHC class II structure. Am. J. Reprod. Immunol. 24, 120–126.
| PubMed |
![](../../media/share/OpenURL_Image.gif)
Zani, M. , Lavitrano, M. , French, D. , Lulli, V. , Maione, B. , Sperandio, S. , and Spadafora, C. (1995). The mechanism of binding of exogenous DNA to sperm cells: factors controlling the DNA uptake. Exp. Cell Res. 217, 57–64.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
![](../../media/share/OpenURL_Image.gif)