TRANSGENIC MICE CONTAINING HUMAN HEMOGLOBIN S

含有人血红蛋白 S 的转基因小鼠

• 批准号: 6241723
• 负责人: FRANK F COSTANTINI
• 金额: $ 22.53万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-04-01 至 1998-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6241723
• 关键词: animal breeding; disease /disorder model; embryonic stem cell; gene mutation; genetically modified animals; hemoglobin Ss; laboratory mouse; model design /development; sickle cell anemia; site directed mutagenesis; tissue mosaicism

While the molecular and cellular properties of hemoglobin S and sickle cells have been studied in detail, the processes leading to vaso- occlusion, painful crisis and other pathophysiological aspects of sickle cell disease remain poorly understood. The ultimate aim of this project is to produce a mouse model of sickle cell disease, which would be of great value for in vivo studies of acute and chronic vaso-occlusive processes, and for developing therapies. The transgenic models that we an others have produced to date generally resemble sickle cell trait, but do not display many of the severe symptoms of homozygous sickle cell disease. One immediate aim is to further characterize the transgenic mouse models we have produced so far, and to breed together different transgenic lines to increase the expression of Hb S (or the variant Hb SAD) and potentially increase the phenotypic effects. However, the success of transgenic approach appears to be limited by the endogenous mouse globins, which combine with human globin chains and dilute the human Hb S. The technology now exists to produce new mouse models in which endogenous hemoglobin is entirely replaced by human Hb S. For this purpose, we will produce targeted mutations in the murine alpha- and beta-globin loci through homologous recombination in embryonic stem (ES) cells, deleting the mouse adult alpha- and beta-globin genes. these mutant globin loci will be introduced into the mouse germ line, where they will be combined (by breeding) with human alpha and human betaS globin transgenes. In an alternative strategy, we will directly replace the mouse adult beta-globin genes with the human betaS gene, and the mouse adult alpha globin genes with the two human adult alpha globin genes, through gene targeting in ES cells. The resulting mice will be characterized to determine the extent to which replacement of mouse Hb with human Hb S results in a model of sickle cell disease.

虽然血红蛋白S和镰刀的分子和细胞特性 细胞已经被详细研究，导致血管- 闭塞、疼痛危象和镰刀的其他病理生理学方面 细胞疾病仍然知之甚少。 该项目的最终目标 是制造一个镰状细胞病的小鼠模型， 对急性和慢性血管闭塞的体内研究具有重要价值 过程以及开发疗法。 我们所研究的转基因模型 到目前为止，其他人已经产生了类似镰状细胞的性状，但 不会表现出许多严重的纯合子镰状细胞病症状 疾病 一个直接的目标是进一步表征转基因 小鼠模型，我们已经生产到目前为止，并繁殖在一起不同的 增加Hb S（或变体Hb SAD）并潜在地增加表型效应。 但 转基因方法的成功似乎受到内源性 小鼠球蛋白，它与人类球蛋白链结合联合收割机， 人血红蛋白S。 这项技术现在已经可以生产新的小鼠模型， 该内源性血红蛋白完全被人血红蛋白S替代。 为此 为了达到这个目的，我们将在鼠α-和 胚胎干细胞（ES）中通过同源重组的β-珠蛋白基因座 细胞，删除小鼠成人α和β-珠蛋白基因。 这些 将突变珠蛋白基因座引入小鼠生殖系，其中 它们将与人类α和人类β相结合（通过繁殖） 珠蛋白转基因 在替代策略中，我们将直接替换 小鼠成体β-珠蛋白基因与人β S基因， 小鼠成人α珠蛋白基因与两个人成人α珠蛋白 基因，通过在ES细胞中的基因靶向。 由此产生的小鼠将是 其特征在于确定小鼠Hb的替代 与人Hb S的接触导致镰状细胞病的模型。