Pig cells that lack the gene for &agr;1-3 galactosyltransferase express low levels of the gal antigen
Pig cells that lack the gene for &agr;1-3 galactosyltransferase express low levels of the gal antigen
复制标题
猪细胞缺乏该基因
作者：
Ajay Sharma;B. Naziruddin;Cunqi Cui;Michael J. Martin;Hui Xu;H. Wan;Y. Lei;C. Harrison;J. Yin;J. Okabe;C. Mathews;A. Stark;C. Adams;J. Houtz;B. S. Wiseman;G. Byrne;J. Logan
背景。灵长类抗体在猪组织上识别的主要抗原是存在于糖脂和糖蛋白上的末端 gal&agr;1-3gal 碳水化合物结构（gal 抗原）。用体细胞生产动物可以使特定基因失活。预计编码&agr;1-3半乳糖基转移酶（合成gal&agr;1-3gal连接的酶）的基因的完全失活将导致猪器官和组织中该抗原的损失，并将在猪到灵长类异种移植中提供生存益处。方法。使用阳性-阴性选择来产生胎猪成纤维细胞，其是&agr;1-3半乳糖基转移酶基因的杂合敲除(+/-)。这些细胞的核移植产生了具有适当基因型的猪胚胎和活产猪。使用来自（+/-）胚胎的细胞的新颖选择方法，我们产生了纯合（−/−）胎猪成纤维细胞。结果。 &agr;1-3 半乳糖基转移酶基因的 Southern 印迹分析表明，我们产生了 (+/-) 猪胚胎、(+/-) 活产猪和 (−/−) 猪胎成纤维细胞。使用一些（但不是全部）小鼠抗 gal 单克隆抗体和致敏人血清进行的荧光激活细胞分选仪 (FACS) 分析显示，(−/−) 细胞仍以对照杂合细胞水平的 1% 至 2% 合成 gal 抗原。结论。敲除&agr;1-3半乳糖基转移酶基因的纯合胎猪成纤维细胞似乎表达低但可检测水平的gal抗原。
Background. The major antigen recognized on pig tissue by primate antibodies is a terminal gal&agr;1-3gal carbohydrate structure (gal antigen) present on glycolipids and glycoproteins. The production of animals from somatic cells allows for the inactivation of specific genes. It is anticipated that the complete inactivation of the gene encoding &agr;1-3 galactosyltransferase, the enzyme that synthesizes the gal&agr;1-3gal linkage, will result in loss of that antigen from pig organs and tissue and will provide a survival benefit in pig-to-primate xenotransplants. Methods. Positive-negative selection was used to produce fetal-pig fibroblasts that were a heterozygous knockout (+/−) of the &agr;1-3 galactosyltransferase gene. Nuclear transfer of these cells generated pig embryos and live born pigs with the appropriate genotype. Using a novel selection method with cells from (+/−) embryos, we produced homozygous (−/−) fetal-pig fibroblast cells. Results. Southern blot analysis of the &agr;1-3 galactosyltransferase gene showed that we had produced (+/−) pig embryos, (+/−) live born pigs, and (−/−) pig-fetal fibroblast cells. Fluorescence-activated cell sorter (FACS) analysis with some, but not all, mouse anti-gal monoclonal antibodies and sensitized human serum showed that (−/−) cells still synthesized the gal antigen at 1 to 2% of the level of control heterozygous cells. Conclusions. Fetal-pig fibroblasts homozygous for the knockout of the &agr;1-3 galactosyltransferase gene appear to express low but detectable levels of the gal antigen.
影响因子：
6.2
作者：
Byrne, G;McCurry, KR;Logan, JS
通讯作者：
Logan, JS