Reproduction, Fertility and Development Reproduction, Fertility and Development Society
Vertebrate reproductive science and technology

Sperm-mediated gene transfer

Marialuisa Lavitrano A B , Marco Busnelli A , Maria Grazia Cerrito A , Roberto Giovannoni A , Stefano Manzini A and Alessia Vargiolu A

A Dipartimento di Scienze Chirurgiche e Terapia Intensiva, Università di Milano, Bicocca, 20052 Milano, Italy.

B Corresponding author. Email:

Reproduction, Fertility and Development 18(2) 19-23
Submitted: 21 September 2005  Accepted: 21 September 2005   Published: 14 December 2005


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.


Brackett B. G.Baranska W.Sawichi W.Koprowski H.1971Uptake of heterologous genome by mammalian spermatozoa and its transfer to ova through fertilization.Proc. Natl Acad. Sci. USA68353357

Brinster R. L.Chen H. Y.Trumbauer M. E.Yagle M. K.Palmiter R. D.1985Factors affecting the efficiency of introducing foreign DNA into mice by microinjecting eggs.Proc. Natl Acad. Sci. USA8244384442

Celebi C.Auvray P.Benvegnu T.Plusquellec D.Jegou B.Guillaudeux T.2002Transient transmission of a transgene in mouse offspring following in vivo transfection of male germ cells.Mol. Reprod. Dev.62477482

Chan P. J.2000Sperm-mediated DNA transfer to cells of the uterus and embryo.Mol. Reprod. Dev.56316318doi:10.1002/(SICI)1098-2795(200006)56:2+<316::AID-MRD23>3.0.CO;2-V

Coffin J. M.1990Molecular mechanism of nucleic acid integration.J. Med. Virol.314349

Francolini M.Lavitrano M.Lamia C. L.French D.Frati L.Cotelli F.Spadafora C.1993Evidence for nuclear internalization of exogenous DNA into mammalian sperm cells.Mol. Reprod. Dev.34133139

Hammer R. E.Pursel V. G.Rexroad C. E.Jr, Wall R. J.Bolt D. J.Ebert K. M.Palmiter R. D.Brinster R. L.1985Production of transgenic rabbits, sheep and pigs by microinjection.Nature315680683doi:10.1038/315680A0

Khoo H. W.Ang L. H.Lim H. B.Wong K. Y.1992Sperm cells as vector for introducing DNA into zebrafish.Aquaculture107119

Kim J. H.Jung-Ha H. S.Lee H. T.Chung K. S.1997Development of a positive method for male stem cell-mediated gene transfer in mouse and pig.Mol. Reprod. Dev.46515526

Kues W. A.Niemann H.2004The contribution of farm animals to human health.Trends Biotechnol.22286294doi:10.1016/J.TIBTECH.2004.04.003

Lavitrano M.Camaioni A.Fazio V. M.Dolci S.Farace M. G.Spadafora C.1989Sperm cells as vectors for introducing foreign DNA into eggs: genetic transformation of mice.Cell57717723doi:10.1016/0092-8674(89)90787-3

Lavitrano M.French D.Zani M.Frati L.Spadafora C.1992The interaction between exogenous DNA and sperm cells.Mol. Reprod. Dev.31161169doi:10.1002/MRD.1080310302

Lavitrano M., Lulli V., Maione B., Sperandio S., and Spadafora C. (1996). Sperm-mediated gene transfer. In ‘Microinjection and Transgenesis. Strategies and Protocols. A Laboratory Manual’. (Eds A. Cid-Arregui and A. Garcìa-Carrancà.) pp. 229–254. (Springer-Verlag: Berlin, Germany.)

Lavitrano M.Forni M.Varzi V.Pucci L.Bacci M. al.1997aSperm-mediated gene transfer: production of pigs transgenic for a human regulator of complement activation.Transplant. Proc.2935083509doi:10.1016/S0041-1345(97)00998-6

Lavitrano M.Maione B.Forte E.Francolini M.Sperandio S.Testi R.Spadafora C.1997bThe interaction of sperm cells with exogenous DNA: a role of CD4 and major histocompatibility complex class II molecules.Exp. Cell Res.2335662doi:10.1006/EXCR.1997.3534

Lavitrano M.Bacci M. L.Forni M.Lazzereschi D.Di Stefano al.2002Production by sperm-mediated gene transfer of human decay accelerating factor (hDAF) transgenic pigs for xenotransplantation.Proc. Natl Acad. Sci. USA9914 23014 235doi:10.1073/PNAS.222550299

Lavitrano M.Forni M.Bacci M. L.Di Stefano C.Varzi V.Wang H.Seren E.2003Sperm mediated gene transfer in pig: selection of donor boars and optimization of DNA uptake.Mol. Reprod. Dev.64284291doi:10.1002/MRD.10230

Magnano A. R.Giordano R.Moscufo N.Baccetti B.Spadafora C.1998Sperm/DNA interaction: integration of foreign DNA sequences in the mouse sperm genome.J. Reprod. Immunol.41187196doi:10.1016/S0165-0378(98)00058-8

Maione B.Lavitrano M.Spadafora C.Kiessling A. A.1998Sperm-mediated gene transfer in mice.Mol. Reprod. Dev.50406409doi:10.1002/(SICI)1098-2795(199808)50:4<406::AID-MRD4>3.0.CO;2-M

Mori T.Guo M. W.Mori E.Shindo Y.Mori N.Fukuda I.Mori T.1990Expression of class II major histocompatibility complex antigen on sperm and its role in fertilization.Am. J. Reprod. Immunol.24910

Muller F.Ivics Z.Erdelyi F.Papp T.Varadi L.Horvath L.Maclean N.1992Introducing foreign genes into fish eggs with electroporated sperm as a carrier.Mol. Mar. Biol. Biotechnol.1276281

Niemann H.Kues W. A.2003Application of transgenesis in livestock for agriculture and biomedicine.Anim. Reprod. Sci.79291317

Rieth A.Pothier F.Sirard M. A.2000Electroporation of bovine spermatozoa to carry DNA containing highly repetitive sequences into oocytes and detection of homologous recombination events.Mol. Reprod. Dev.57338345doi:10.1002/1098-2795(200012)57:4<338::AID-MRD5>3.0.CO;2-K

Rottmann O. J.Antes R.Hoefer P.Maierhofer G.1991Liposomes mediate gene transfer via spermatozoa into avian egg cells.J. Anim. Breed. Genet.1096470

Schellander K.Peli J.Schmall F.Brem G.1995Artificial insemination in cattle with DNA-treated sperm.Anim. Biotech.64150

Sciamanna I.Piccoli S.Barberi L.Zaccagnini G.Magnano A. R.Giordano R.Campedelli P.Hodgson C.Lorenzini R.Spadafora C.2000DNA dose and sequence dependence in sperm-mediated gene transfer.Mol. Reprod. Dev.56301305

Shemesh M.Gurevich M.Harel-Markowitz E.Benvenisti L.Shore L. S.Stram Y.2000Gene integration into bovine sperm genome and its expression in transgenic offspring.Mol. Reprod. Dev.56Suppl. 2306308doi:10.1002/(SICI)1098-2795(200006)56:2+<306::AID-MRD21>3.0.CO;2-3

Smith K.Spadafora C.2005Sperm-mediated gene transfer: applications and implications.Bioessays27551562doi:10.1002/BIES.20211

Spadafora C.1998Sperm cells and foreign DNA: a controversial relation.BioEssays20955964doi:10.1002/(SICI)1521-1878(199811)20:11<955::AID-BIES11>3.0.CO;2-8

Sperandio S.Lulli V.Bacci M. L.Forni M.Maione B.Spadafora C.Lavitrano M.1996Sperm mediated DNA transfer in bovine and swine species.Anim. Biotechnol.75977

Tsai H. J.Lai C. H.Yang H. S.1997Sperm as a carrier to introduce an exogenous DNA fragment into the oocyte of Japanese abalone (Haliotis divorsicolor suportexta).Transgenic Res.68595

Umemoto Y.Sasaki S.Kojima Y.Kubota H.Kaneko T.Hayashi Y.Kohri K.2005Gene transfer to mouse testes by electroporation and its influence on spermatogenesis.J. Androl.26264271

Wall R. J.1999Sperm-mediated gene transfer: advances in sperm cell research and applications.Transgenic Res.8313315

Wall R. J.2001Pronuclear microinjection.Cloning Stem Cells3209220doi:10.1089/15362300152725936

Wall R. J.2002New gene transfer methods.Theriogenology57189201doi:10.1016/S0093-691X(01)00666-5

Webster N. L.Forni M.Bacci M. L.Giovannoni R.Razzini al.2005Multi-transgenic pigs expressing three fluorescent proteins produced with high efficiency by sperm mediated gene transfer.Mol. Reprod. Dev.726876doi:10.1002/MRD.20316

Wu G. M.Nose K.Mori E.Mori T.1990Binding of foreign DNA to mouse sperm mediated by its MHC class II structure.Am. J. Reprod. Immunol.24120126

Zani M.Lavitrano M.French D.Lulli V.Maione B.Sperandio S.Spadafora C.1995The mechanism of binding of exogenous DNA to sperm cells: factors controlling the DNA uptake.Exp. Cell Res.2175764

Full Text PDF (170 KB) Export Citation