Harrison Sharon J, Guidolin Angelo, Faast Renate, Crocker Lesley A, Giannakis Chris, D'Apice Anthony J F, Nottle Mark B, Lyons Ian
Reproduction and Biotechnology Unit, BresaGen Limited, Adelaide, South Australia, Australia.
Transgenic Res. 2002 Apr;11(2):143-50. doi: 10.1023/a:1015262108526.
Pigs are currently considered the most likely source of organs for human xenotransplantation because of anatomical and physiological similarities to humans, and the relative ease with which they can be bred in large numbers. A severe form of rejection known as hyperacute rejection has been the major barrier to the use of xenografts. Generating transgenic pigs for organ transplantation is likely to involve precise genetic manipulation to ablate the alpha(1,3) galactosyltransferase (galT) gene. In contrast to the mouse, homologous recombination in livestock species to ablate genes is hampered by the inability to isolate functional embryonic stem cells. However, nuclear transfer using genetically targeted cultured somatic cells provides an alternative means to producing pigs deficient for galT. In this study we successfully produced galT+/- somatic porcine fetal fibroblasts using two approaches; positive negative selection (PNS) using an isogenic targeting construct, and with a promoterless vector using non-isogenic DNA.
由于猪在解剖学和生理学上与人类相似，并且相对容易大量繁殖，目前猪被认为是人类异种移植器官的最可能来源。一种被称为超急性排斥的严重排斥形式一直是使用异种移植物的主要障碍。生成用于器官移植的转基因猪可能需要精确的基因操作来消除α（1,3）半乳糖基转移酶（galT）基因。与小鼠不同，家畜物种中用于基因消除的同源重组因无法分离功能性胚胎干细胞而受到阻碍。然而，使用基因靶向培养的体细胞进行核移植提供了一种生产缺乏galT的猪的替代方法。在本研究中，我们使用两种方法成功地产生了galT+/- 体细胞猪胎儿成纤维细胞；使用同基因靶向构建体的正负选择（PNS），以及使用非同源DNA的无启动子载体。
