Lavitrano Marialuisa, Busnelli Marco, Cerrito Maria Grazia, Giovannoni Roberto, Manzini Stefano, Vargiolu Alessia
Dipartimento di Scienze Chirurgiche e Terapia Intensiva, Università di Milano, Bicocca, 20052 Milano, Italy.
Reprod Fertil Dev. 2006;18(1-2):19-23. doi: 10.1071/rd05124.
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.
自1989年以来，一种生产转基因动物的新方法问世了，即精子介导的基因转移（SMGT），其基于精子细胞结合并内化外源DNA分子以及在受精时将它们转移到卵母细胞中的内在能力。我们首先在一个小动物模型中描述了SMGT程序，在小鼠中报道了高效率。此外，我们成功地对该技术进行了调整和优化，使其适用于大型动物；事实上，它在生成人衰变加速因子转基因猪系以及导入了三种不同报告基因（即增强型绿色荧光蛋白、增强型蓝色荧光蛋白和红色荧光蛋白）的多基因转基因猪方面效率很高。与其他方法相比，发现SMGT技术的主要优点是其高效率、低成本和易于使用。此外，SMGT不需要处理胚胎或昂贵的设备。精子介导的基因转移还可用于生成多基因转基因猪，这作为医学研究、农业和制药应用，特别是异种移植的大型动物模型将是有益的，异种移植需要对供体猪进行广泛的基因操作，使其适合移植给人类。
