A TRANSGENIC PIG MODEL FOR SICKLE CELL DISEASE

我们已经开始制作用于镰刀的转基因猪模型 细胞病(SS病)为镰刀研究提供便利 细胞病理学与新药和基因的评价 治疗。之前试图制造转基因模型 用鼠标都没能产生临床上的 SS病的症状学，尽管产生了， 在某些情况下，大量的人类镰刀 血红蛋白(HBs)和细胞内的证据 血红蛋白的聚合。美国政府的失败 转基因小鼠将患上严重的镰状细胞疾病 病理似乎是由于血管的不同。 老鼠和人类之间的解剖学和生理学。论 另一方面，猪为人类提供了一个很好的模型 循环生理学，重要的是，就我们的目的而言， 表现出对血管活性化合物的同源反应， 毛细血管床密度、脑血管 疾病和某些红细胞结构特性。这个 生产转基因农场(全尺寸)猪的技术是 也是久负盛名。 生产高水平珠蛋白转基因动物 转基因表达，一种被称为 β-珠蛋白基因座控制区(LCR)，通常位于 在β-珠蛋白基因簇的上游，已经被证明 在CI中是必需的。我们已经准备好DNA结构 含有人类β-珠蛋白的功能部分 与人类α-2珠蛋白基因连锁的LCR(微型LCR)或 一种修饰的人类镰状β-珠蛋白基因 促进聚合的突变(β-SAD，包含 Antilles和D Punjab)的其他突变 对猪进行微量注射。同时，为了 便于维护和临床分析 我们已经开始开发NIH转基因猪--小型猪 作为转基因系统。使用纯化的卵泡刺激素方案， 已获得显著的超数排卵(40) 蛋/猪)。在繁殖之后，受精卵已经被 康复和显微注射手术程序 再植入和维持妊娠正在进行中 开发将转基因技术扩展到更多 大小适中的猪品种。一旦建立起来， 产生高水平人类的转基因猪模型 镰刀状的血红蛋白和SS病的病理， 将极大地加速我们理解和理解 治疗这种疾病。

We have begun to make a transgenic pig model for sickle cell disease (SS disease) to facilitate studies for sickle cell pathology and evaluations of new drug and gene therapies. Previous attempts to make transgenic models using the mouse have failed to produce the clinical symptomatology of SS disease, in spite of the production, in some cases, of significant amounts of human sickle hemoglobin (HbS) and evidence for intracellular polymerization of hemoglobin. The failure of the transgenic mice to develop significant sickle cell disease pathology appears to be due to differences in the vascular anatomy and physiology between mice and humans. On the other hand, the pig provides an excellent model for human circulatory physiology, significantly, for our purposes, evincing cognate responses to vaso-active compounds, similarities in capillary bed density, cerebrovascular diseases and certain red cell structural properties. The technology to produce transgenic farm (full-size) pigs is also well established. To produce transgenic animals with high level globin transgene expression, a genetic regulatory region called by beta-globin locus control region (LCR), usually located upstream of the beta-globin gene cluster, has been shown to be required in cis. We have prepared DNA constructions containing the functional portions of the human beta-globin LCR (mini-LCR) linked to the human alpha-2-globin gene or a modified human sickle beta-globin gene with additional mutations to promote polymerization (beta SAD, containing additional mutations Antilles and D Punjab) for microinjection into the pigs. At the same time, to facilitate the maintenance and clinical analysis of the transgenic pigs we have begun to develop the NIH--minipig as a transgenic system. Using a regimen of purified FSH, significant superovulation has been obtained (40) eggs/pig). Following breeding, fertilize eggs have been recovered and procedures for microinjection, surgical reimplantation and maintenance of pregnancy are being developed to extend the transgenic technology to this more conveniently-sized pig variety. Once established, the transgenic pig model that produces high levels of human sickle hemoglobin and demonstrates SS disease pathology, would greatly accelerate our efforts to understand and treat the disease.