Rubin E M, Witkowska H E, Spangler E, Curtin P, Lubin B H, Mohandas N, Clift S M
Division of Cell and Molecular Biology, Lawrence Berkeley Laboratory, University of California, Berkeley 94720.
J Clin Invest. 1991 Feb;87(2):639-47. doi: 10.1172/JCI115041.
To develop an animal model for sickle cell anemia, we have created transgenic mice that express a severe naturally occurring human sickling hemoglobin, Hb S Antilles. Due to its low solubility and oxygen affinity, Hb S Antilles has a greater propensity to cause red cell sickling than Hb S. To make transgenic animals that express a high level of Hb S Antilles, the erythroid-specific DNAse I hypersensitive site II from the human beta-globin cluster was linked independently to the human alpha 2-globin gene and to the beta S Antilles gene. Embryos were injected with both constructs simultaneously and seven transgenic mice were obtained, three of which contained both the human alpha and the human beta S Antilles transgene. After crossing the human transgenes into the mouse beta-thalassemic background a transgenic mouse line was derived in which approximately half the beta-globin chains in the murine red cells were human beta S Antilles. Deoxygenation of the transgenic red cells in vitro resulted in extensive sickling. An increase of in vivo sickling was achieved by placing these transgenic mice in a low oxygen environment. This murine model for red cell sickling should help to advance our understanding of sickle cell disease and may provide a model to test therapeutic interventions.
为了建立镰状细胞贫血的动物模型，我们培育了转基因小鼠，使其表达一种严重的天然存在的人类镰状血红蛋白——Hb S安的列斯型。由于其低溶解性和氧亲和力，Hb S安的列斯型比Hb S更易导致红细胞镰变。为了培育出能高水平表达Hb S安的列斯型的转基因动物，将来自人类β-珠蛋白基因簇的红系特异性DNA酶I超敏位点II分别与人类α2-珠蛋白基因和βS安的列斯型基因连接。同时用这两种构建体对胚胎进行注射，获得了7只转基因小鼠，其中3只同时含有人类α和人类βS安的列斯型转基因。将人类转基因导入小鼠β地中海贫血背景后，得到了一个转基因小鼠品系，其中小鼠红细胞中约一半的β-珠蛋白链是人类βS安的列斯型。体外转基因红细胞的脱氧导致广泛的镰变。将这些转基因小鼠置于低氧环境中可增加体内镰变。这种红细胞镰变的小鼠模型应有助于增进我们对镰状细胞病的理解，并可能为测试治疗干预措施提供一个模型。
