Sperm-mediated gene transfer.
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.
Lavitrano
Marialuisa
M
Dipartimento di Scienze Chirurgiche e Terapia Intensiva, Università di Milano, Bicocca, 20052 Milano, Italy. marialuisa.lavitrano@unimib.it
Busnelli
Marco
M
Cerrito
Maria Grazia
MG
Giovannoni
Roberto
R
Manzini
Stefano
S
Vargiolu
Alessia
A
D000818:Animals; D030801:Animals, Genetically Modified; D018014:Gene Transfer Techniques; D008297:Male; D013094:Spermatozoa / Q000502:physiology*; D013552:Swine / Q000235:genetics*