Production of pig with low immune response for human by gene transfection

目的：在器官移植中，供体短缺是世界性的难题.为了解决器官短缺问题，人们考虑进行异种移植。在猪-人异种器官移植中，人异种天然抗体和补体引起的超急性排斥反应是需要克服的一大障碍。本研究尝试用天然抗原（1，3-半乳糖基转移酶）基因敲除猪，研究1，3-半乳糖基转移酶反义RNA载体是否能减少主要异种抗原，以控制超急性排斥反应。方法：构建1，3-半乳糖基转移酶第9外显子基因敲除载体和α-1，3-半乳糖基转移酶反义RNA载体。并转染到猪类干细胞中，分析了细胞的猪异种抗原。结果：基因敲除载体转染干细胞样细胞后，1，3-半乳糖不能降解。虽然反义核酸是瞬时表达的，但它能够降低猪干细胞样细胞的异种抗原。结论：通过转染表达反义RNA并阻止α-1，3-半乳糖基转移酶蛋白翻译的方法有可能成为控制超急性排斥反应的手段之一。

Objective: In organ transplantation, shortage of donor is a big problem all over the -world. For the solution of the organ shortage xenotransplantation is considered. In pig-to-human xenotransplantation, the hyper-acute rejection triggered by human xenoreactive natural antibody and complement is a big barrier we should overcome. In this study we tried to make knock-out pig in which natural antigen (a 1,3-galactose) and studied whether the antisense RNA vector for a 1,3 -galactosyltransferase could decrease the major xenoantigen in order to control the hyperacute rejection. Methods: We created both knock out vector of 1,3-galactosyltransferase in exon 9 and the antisense of alpha-i, 3-galactosyltransferase RNA vector. And we transfected to a pig stem like cell and analyzed the porcine xenoantigen of the cells. Result: Knock out vector was tranfected into the stem like cells but a 1,3-galactose could not be deceased. Although the antisense was transient expression, it was able to decrease the xenoantigen of a pig stem like cell.. Conclusion: The method that expressed an antisense RNA by transfection and prevented translation of alpha-1, 3-galactosyltransferase protein has a possibility that would be one of the means to control hyperacute rejection.