ANALYSIS OF DIRECT ACTIONS OF TRANSFORMING PROTEINS OF ONCOGENIC VIRUSES

我们试图开发一种方法， 克隆基因到造血干细胞中。 造血干细胞 选择这些细胞是因为（A）这些细胞是多能的，（B）可以在 允许对单个子代进行体外分析，和（C）将重构 受到致命辐射的老鼠的造血区。 后一 这使得在体内和体内进行长期调节的研究成为可能。 体外，也开辟了新的可能性，基因治疗， 镰状细胞性贫血和地中海贫血等造血系统疾病。 我们有 从两个方面着手进行这些实验。 首先，我们获得了 在制备含有以下基因的病毒构建体方面的丰富经验： 感兴趣和适当调节序列。 我们现在已经完成了 这表明含有EJ膀胱癌的结构 基因将感染并转化红系祖细胞， 提供生长优势而不阻碍分化。 为 红系细胞成熟并合成血红蛋白和其它红细胞 它们也合成大量的癌基因蛋白 第21页。 第二，我们使用类似的结构来证明第一个 多能造血干细胞可以直接感染的时间， 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.