ERYTHROID DIFFERENTIATION IN FRIEND LEUKEMIA CELLS

友人白血病细胞中的红细胞分化

• 批准号: 2086615
• 负责人: David Housman
• 金额: $ 25万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 1978
• 资助国家: 美国
• 起止时间: 1978-05-01 至 1996-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2086615
• 关键词: DNA binding protein; Friend leukemia; R factors; affinity chromatography; cell differentiation; cell transformation; drug resistance; erythroid stem cell; erythroleukemia; erythropoiesis; gel electrophoresis; gene expression; genetic manipulation; genetic transcription; globin; hematopoiesis; hematopoietic stem cells; laboratory mouse; messenger RNA; molecular cloning; tissue /cell culture; transfection /expression vector

The overall objective of this proposal is to understand the processes which control hematopoietic differentiation at the molecular level. The following specific aims will be addressed: 1. To develop a more complete understanding of the factors which control globin gene expression by: a.) Evaluating the impact on the expression of the mouse beta globin gene of mutation at sites in the IVS2 region which exhibit specific binding and footprinting for tissue specific binding proteins B1 and B2 b.) Further characterizing and purifying DNA binding proteins B1 and B2 and c.) Extending DNA binding studies to additional sites in the mouse beta globin gene. 2. To extend our initial studies identifying mRNA regions which contribute to mRNA stability in beta globin and c-fos mRNAs by: a.) Developing additional chimeric constructs between human c- fos and beta globin mRNAs to discriminate with greater precision mRNA sequences or structures which contribute to the differences in stability between these mRNA species. b.) Extending the chimeric mRNA approach to additional mRNA species including alpha globin, c-myc and delta globin mRNAs. C.) Identifying an inducible expression system which will allow us to extend the evaluation of mRNA stability to differentiating hematopoietic cells d.) Developing biochemical and genetic methods for identifying cellular mechanisms which respond to signals within mRNA molecules which lead to differences in half life. 3.) To develop somatic cell genetic approaches to hematopoietic differentiation by: a. Applying gene transfer procedures to determine the significance of changes in expression levels of specific polypeptides in the control of the differentiation program. b. Developing the use of retroviral vectors for insertional mutagenesis in the MEL cell system c. Developing multidrug resistance gene system for use as a selection system for bone marrow progenitor cells and pluripotent stem cells.

该提案的总体目标是了解 控制造血分化的过程 分子水平。 将实现以下具体目标： 1. 对因素有更全面的了解 它通过以下方式控制珠蛋白基因表达： a.) 评估 对小鼠β珠蛋白基因表达的影响 IVS2 区域中表现出特异性结合的位点发生突变 以及组织特异性结合蛋白 B1 和 B2 的足迹 b.) 进一步表征和纯化 DNA 结合蛋白 B1 和 B2 和 c.) 将 DNA 结合研究扩展到其他位点 小鼠β珠蛋白基因。 2. 扩展我们的初步研究，确定 mRNA 区域 通过以下方式促进 β 珠蛋白和 c-fos mRNA 的 mRNA 稳定性： a.) 开发人类 c-之间的额外嵌合构建体 fos 和 beta 珠蛋白 mRNA 可以更精确地进行区分 mRNA序列或结构有助于 这些 mRNA 种类之间的稳定性差异。 b.) 将嵌合 mRNA 方法扩展到其他 mRNA 种，包括 α 珠蛋白、c-myc 和 delta 珠蛋白 mRNA。 C.) 识别诱导表达系统，这将使我们能够 将 mRNA 稳定性的评估扩展到分化 造血细胞 d.) 发育生化和遗传 识别响应的细胞机制的方法 mRNA 分子内的信号导致一半的差异 生活。 3.) 开发体细胞遗传学方法 造血分化通过：a．应用基因转移 确定表达变化意义的程序 控制分化的特定多肽水平 程序。 b.开发逆转录病毒载体的用途 MEL 细胞系统中的插入突变 c．发展 多药耐药基因系统用作选择系统 骨髓祖细胞和多能干细胞。