PRODUCTION OF Alpha-1,3-GALACTOSYLTRANSFERASE DEFICIENT

用于移植的人类捐献器官的持续严重短缺导致人们对使用猪作为器官捐献产生了极大的兴趣。猪器官的超急性排斥反应，由预先形成的灵长类动物抗α -1,3-半乳糖表位抗体介导，是最终使用猪作为人类移植器官供体必须克服的障碍。从猪细胞中去除1,3-半乳糖表位是解决超急性排斥反应问题的一种创新和必要的方法，因为其他方法已被证明不足以消除关键器官损伤或只有短暂的有效性。BioTransplant计划生产用于实体器官异种移植的完全缺乏α -1,3-半乳糖表位表达的猪。这将通过使用携带α - 1,3-半乳糖转移酶（galtransferase）基因敲除位点的供体核质体的核转移技术来实现。由于缺乏生产具有基因靶向改变的转基因大型动物的替代方法，猪核移植的发展是必要的。建议的商业应用：器官移植是终末期器官疾病的一种成熟的治疗方法。然而，全球器官严重短缺，潜在器官移植接受者的等待名单越来越长。最近的统计数据表明，仅在美国就有6万多名终末期器官疾病患者等待器官移植，每年约有4000人在等待过程中死亡。异种器官移植是解决供体器官长期短缺的唯一可能的长期解决方案。

Persistent severe shortages of human donor organs for transplantation have resulted in great interest in the use of the pig as an organ donor. Hyperacute rejection of pig organs, mediated by preformed primate antibodies against alpha-1,3-galactosyl epitopes, is a barrier that must be overcome for eventual use of pigs as organ donors for human transplants. Elimination of 1,3- galactosyl epitopes from pig cells is an innovative and necessary approach to the problem of hyperacute rejection, as other approaches have proven insufficient to eliminate critical organ damage or to have only transient effectiveness. BioTransplant proposes to produce pigs, for use in solid organ xenotransplantation, that are totally deficient in the expression of alpha-1,3- galactosyl epitopes. This will be accomplished through use of nuclear transfer technologies with donor karyoplasts carrying a genetic knockout of the alpha- 1,3-galactosyltransferase (galtransferase) locus. Development of nuclear transfer in pigs is necessitated by the lack of alternative methods for production of transgenic large animals with gene-targeted alterations. PROPOSED COMMERCIAL APPLICATIONS: Organ transplantation is an established therapy for end-stage organ disease. However, there is a critical worldwide shortage of organs and increasingly large waiting lists have been established for potential organ transplant recipients. Recent statistics indicate that more than 60,000 people with end stage organ disease in the United States alone await organ transplants and that about 4,000 people die annually while waiting. Xenotransplantation is the only likely long-term solution to the chronic shortage of donor organs.