IMPROVE TRANSGENIC TECHNOLOGY--USE OF MALE GERM CELLS

描述(改编自申请人的摘要)：转基因技术， 涉及从哺乳动物身上增加或删除基因的 基因组，已被证明是最有价值的工具之一 遗传学。转基因动物对人类的知识至关重要 遗传病、发育规划和遗传相互作用。 转基因也被用于家畜的改良和 药品生产。转基因动物的产生 将转基因微量注射到受精卵中是可重复性的， 没有必要改进已经在发挥作用的东西。然而，a 这种转基因技术的重大缺点是它依赖于 充足的受精卵供应。在灵长类等较大的动物中， 猪和牛，生产500个或更多鸡蛋的成本(10-20个鸡蛋或 少/动物)贵得令人望而却步。然而，在许多情况下 转基因灵长类动物将是更适合研究人类疾病的模型 用来测试疗法。大型家畜会更有效地 治疗药物和异种移植的商业生产。 由于睾丸含有丰富的生殖细胞[例如，0.5x 10(8)/小鼠睾丸]，申请者提议产生转基因动物 使用雄性生殖细胞。转基因动物将由两个 不同的方法：1)将基因直接输送到体内的雄性生殖细胞 使用病毒载体；以及2)将从睾丸中获取生殖细胞， 在体外导入DNA，并将稳定的细胞移植到 经化学处理或照射的不含内源性细菌的睾丸 细胞。转基因的生殖细胞将重新填充到睾丸和 动物将通过自然交配产生活的转基因后代。 由这两种技术中的任何一种产生的转基因动物很可能是 效率高、性价比高，而且需要的动物要少得多。

DESCRIPTION (Adapted from the applicant's abstract): Transgenic technology, involving either the addition or deletion of a gene from the mammalian genome, has proven to be one of the most valuable tools for the study of genetics. Transgenic animals have been crucial to the knowledge of human genetic diseases, developmental programming, and genetic interactions. Transgenesis has also been used for the improvement of livestock and for the production of pharmaceuticals. The generation of transgenic animals by microinjection of the transgene into fertilized eggs is reproducible and there has been no necessity to improve what is already working. However, a significant drawback of this transgenic technique is that it relies on a plentiful supply of fertilized eggs. In larger animals such as primates, pigs, and cows, the cost of producing 500 or more eggs (at 10-20 eggs or less/animal) is prohibitively expensive. Yet there are many instances when transgenic primates would be a more appropriate model for human diseases and for testing therapies. Large domestic animals would be more effective for the commercial production of therapeutic drugs and for xenotransplantation. Since the testis contains an abundance of germ cells [e.g., 0.5 x 10(8)/mouse testis], the applicants propose to generate transgenic animals using male germ cells. Transgenic animals will be generated by two different methods: 1) direct delivery of genes to male germ cells in vivo using viral vectors; and 2) germ cells will be harvested from the testis, transfected with DNA in vitro, and stable cells transplanted into either chemically-treated or irradiated testes that contain no endogenous germ cells. The transfected germ cells would repopulate the testis and the animals would produce live transgenic offspring by natural mating. Transgenic animals produced by either of these two techniques will likely be efficient, cost-effective, and will require far fewer animals.