IMPROVE TRANSGENIC TECHNOLOGY--USE OF MALE GERM CELLS

改进转基因技术——利用雄性生殖细胞

• 批准号: 2751040
• 负责人: CAROL W. READHEAD
• 金额: $ 14.78万
• 依托单位: CEDARS-SINAI MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-08-01 至 1999-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2751040
• 关键词: biotechnology; genetic manipulation; genetic techniques; genetically modified animals; germ cells; laboratory mouse; male; sperm; technology /technique development; testis; transfection; transfection /expression vector

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.

描述（改编自申请人摘要）：转基因技术， 涉及从哺乳动物中添加或缺失基因 基因组，已被证明是研究的最有价值的工具之一， 遗传学 转基因动物对人类认识 遗传疾病、发育规划和遗传相互作用。 转基因也已用于改良家畜和用于 药品生产。 转基因动物的产生 将转基因显微注射到受精卵中是可重复的， 没有必要改进已经起作用的东西。 但 这种转基因技术的显著缺点是它依赖于 丰富的受精卵供应。 在大型动物如灵长类中， 猪和牛，生产500个或更多鸡蛋的成本（10-20个鸡蛋或 更少/动物）是非常昂贵的。 然而，有许多情况下， 转基因灵长类动物将是人类疾病更合适的模型， 用于测试疗法。 大型家畜将更有效地为 治疗药物的商业化生产和异种移植。 由于睾丸含有丰富的生殖细胞[例如，0.5 X 10（8）/小鼠睾丸]，申请人提出产生转基因动物 使用男性生殖细胞。 转基因动物将由两个 不同的方法：1）将基因直接递送到体内雄性生殖细胞 使用病毒载体;和2）从睾丸收获生殖细胞， 在体外用DNA转染，并将稳定的细胞移植到 不含内源性细菌的化学处理或辐照睾丸 细胞 转染的生殖细胞将重新填充睾丸， 动物可以通过自然交配产生活的转基因后代。 通过这两种技术中的任何一种生产的转基因动物都可能是 高效、成本效益高，而且需要的动物要少得多。