Transgenic Goats Produced by Germ Cell Transplantation

生殖细胞移植产生的转基因山羊

• 批准号: 6933699
• 负责人: INA DOBRINSKI
• 金额: $ 37.43万
• 依托单位: GTC BIOTHERAPEUTICS, INC.
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-04-01 至 2007-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6933699
• 关键词: adeno associated virus group; biotherapeutic agent; cell transplantation; gene expression; genetically modified animals; germ cells; goats; in vitro fertilization; mammary gland; milk; recombinant proteins; somatotropin; sperm; technology /technique development; tissue /cell culture; transfection /expression vector

DESCRIPTION (provided by applicant): The objective of this proposal is to produce transgenic goats by manipulation of the male germ line through male germ cell transfection and transplantation. The goats will carry the gene for human growth hormone (rhGH) under the control of a mammary gland specific promoter such that the milk of transgenic females will be an efficient source of protein for purification of human growth hormone. The milk of transgenic goats is a safe, abundant and cost effective production source for recombinant human biopharmaceuticals. GTC Biotherapeutics (formerly Genzyme Transgenics Corp.) has nearly two dozen commercial transgenic programs in development. However, even state-of-the-art methods for generating transgenic founder (first generation) goats remain relatively inefficient and expensive. Methods have been developed in rodents for transplanting genetically modified male germ cells into recipient males and mating these animals to produce transgenic offspring. During phase I of this project we successfully adapted the systems of germ cell transfection and transplantation to goats and demonstrated feasibility that recipient goats produce offspring carrying the genetic makeup of the germ cell donor animal. Now we propose to: 1) produce goats making transgenic sperm after transplantation of transfected germ cells; 2) generate transgenic embryos by in vitro fertilization and produce live transgenic offspring carrying expressing a commercially significant protein in their milk; and 3) establish a system for caprine germ cell culture to make the approach universally applicable to a wider range of transgene constructs. Successful introduction of a transgene through the male germ line will increase the efficiency and reduce the cost of generating founder animals, which is a critical early step in the transgenic production process.

描述（由申请人提供）：本提案的目的是通过雄性生殖细胞转染和移植操作雄性生殖系来生产转基因山羊。山羊将在乳腺特异性启动子的控制下携带人类生长激素（rhGH）基因，这样转基因雌性山羊的乳汁将成为纯化人类生长激素的有效蛋白质来源。转基因山羊奶是重组人类生物制药的安全、丰富且具有成本效益的生产来源。 GTC Biotherapeutics（原 Genzyme Transgenics Corp.）有近两打商业转基因项目正在开发中。然而，即使是产生转基因创始人（第一代）山羊的最先进方法仍然相对低效且昂贵。已经在啮齿类动物中开发出将转基因雄性生殖细胞移植到受体雄性体内并使这些动物交配以产生转基因后代的方法。在该项目的第一阶段，我们成功地将生殖细胞转染和移植系统应用于山羊，并证明了受体山羊产生携带生殖细胞供体动物基因组成的后代的可行性。现在我们建议：1）通过移植转染的生殖细胞来生产产生转基因精子的山羊； 2) 通过体外受精产生转基因胚胎，并产生在乳汁中表达具有商业意义的蛋白质的活转基因后代； 3）建立山羊生殖细胞培养系统，使该方法普遍适用于更广泛的转基因构建体。通过雄性种系成功引入转基因将提高生产起始动物的效率并降低其成本，这是转基因生产过程中关键的早期步骤。