ANALYSIS OF DIRECT ACTIONS OF TRANSFORMING PROTEINS OF ONCOGENIC VIRUSES

我们试图开发一种方法，使转移成为可能 克隆的基因进入了造血干细胞。造血干细胞是 选择这些细胞是因为(A)这些细胞是多潜能的，(B)可以在 个体后代的体外许可分析，以及(C)将重建 受到致命照射的小鼠的造血室。后者 属性使体内和体内长期调控研究成为可能 并为此类疾病的基因治疗开辟了新的可能性 如镰状细胞性贫血和地中海贫血等血液系统疾病。我们有 从两个方面探讨了这些实验。第一，我们取得了 在制造含有病毒基因的病毒载体方面有丰富的经验 兴趣和适当的调控序列。我们现在已经完成了工作 这表明含有EJ膀胱癌的结构 基因会感染和转化红系祖细胞 在不阻碍差异化的情况下提供增长优势。作为 红系细胞成熟并合成血红蛋白和其他红细胞 蛋白质，它们还合成大量的癌基因蛋白质。 第21页。其次，我们已经使用类似的构造来演示第一个 多能造血干细胞直接感染的时间和 它们分化的后代将表达转移的基因 信息。

We have attempted to develop the methodology which will allow the transfer of cloned genes into hemopoietic stem cells. Hemopoietic stem cells were chosen because (A) these cells are multipotential, (B) can be cultured in vitro permitting analysis of individual progeny, and (C) will reconstitute the hemopoietic compartment of a lethally irradiated mouse. This latter propery makes feasible the study of long-term regulation in vivo and in vitro and also opens new possibilities for genetic therapy of such hemopoietic diseases as sickle cell anemia and thalassemia. We have approached these experiments from two directions. First, we have gained substantial experience with making viral constructs containing genes of interest and appropriate regulatory sequences. We have now completed work which demonstrates that constructions containing the EJ bladder carcinoma gene will infect and transform erythroid progenitors in a manner that provides a growth advantage without blocking differentiation. As the erythroid cells mature and synthesize hemoglobin and other red cell proteins, they also synthesize copious amounts of the oncogene protein p21. Second, we have used similar constructs to demonstrate for the first time that multipotent hemopoietic stem cells can be directly infected and that their differentiated progeny will express the transferred genetic information.