MOLECULAR BIOLOGY OF LYMPHOCYTE AND NEURONAL GROWTH

淋巴细胞和神经元生长的分子生物学

• 批准号: 2022193
• 负责人: DAVID BALTIMORE
• 金额: $ 38.28万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-12-01 至 2001-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2022193
• 关键词: developmental genetics; developmental neurobiology; gene rearrangement; genetically modified animals; immunogenetics; immunoglobulin genes; laboratory mouse; leukocyte activation /transformation; lymphocyte; molecular biology; neurogenesis; neurons; oligonucleotides; tissue /cell culture; transcription factor

DESCRIPTION: The overall aim of this program is to gain as complete a knowledge as possible of the events that transpire during the differentiation of B lymphocytes and to begin to extend the analysis to the nervous system. This work is particularly relevant to the study of autoimmune disease, immunologic responses, immunologic memory and neuronal deficits. The specific aims of the next granting period are to understand the role of a single transcriptional enhancer, the intron enhancer of the Kappa (K) light chain; to understand the immunoglobulin gene rearrangement process and its control at a biochemical level; to develop a method to mark memory cells and to extend the methods of analysis of immunodifferentiation to the study of neuronal differentiation. Kappa gene expression is thought to be controlled by 2 enhancers. The various roles of these enhancers in immunodifferentiation will be analyzed using homologous recombination to make critical mutants in ES cells that can then be used to reconstitute the lymphoid systems of RAG-2-deleted mice. ES cell homologous recombination will also be used to insert K regulatory elements into the l genes to examine whether the K/l light chain ratio in the mouse is a consequence of the relative strengths of the transcriptional regulatory elements of the two light chain genes. The immunoglobulin gene rearrangement process is a unique process of DNA rearrangement. To study it, the various relevant proteins will be purified and their in vitro activities will be examined. This analysis will be extended to chromatin and methylated DNA to attempt to understand the various levels of control of the rearrangement process. Analysis in cells will be used to uncover more complex levels of control. There is no unambiguous marker on the surface of memory B and T cells. Transgenic mice will be generated in which the memory B and T cells will be genetically marked and thus can be identified and examined unambiguously. The study of immunodifferentiation has benefitted from the availability of clonal cell lines and methods of chimeric analysis. To extend these benefits to the study of neuronal differentiation, methods will be developed to prepare clonal derivatives of differentiated neurons using oncogenes and a precise labeling procedure. Also to study the role of otherwise lethal genes in neuronal development, a method of chimeric analysis will be developed. It will be applied particularly to analyzing the role of the NF-KB transcription factor in nervous system signaling.

描述：该计划的总体目标是获得完整的 尽可能地了解事件发生期间发生的事件， B淋巴细胞的分化，并开始将分析扩展到 神经系统 这项工作特别与研究 自身免疫性疾病，免疫应答，免疫记忆和神经元 赤字 下一个授予期的具体目标是了解 一个单一的转录增强子，内含子增强子的作用， κ（K）轻链;了解免疫球蛋白基因重排 过程及其在生化水平上的控制;开发一种方法来标记 记忆细胞和扩展免疫分化分析方法 神经元分化的研究。 Kappa基因表达被认为是 由2个增强子控制。 这些增强子的各种作用， 免疫分化将使用同源重组进行分析， 在胚胎干细胞中制造关键突变体，然后可以用来重建 RAG-2缺失小鼠的淋巴系统。 ES细胞同源重组 也将用于将K调节元件插入到I基因中， 检查小鼠中的K/l轻链比率是否是 这两个转录调控元件的相对强度 轻链基因 免疫球蛋白基因重排过程是一个 独特的DNA重排过程 为了研究它，各种相关的 蛋白质将被纯化，并且它们的体外活性将被检测。 这项分析将扩展到染色质和甲基化DNA，试图 了解重排过程的各个控制级别。 细胞分析将用于揭示更复杂的控制水平。 在记忆B和T细胞的表面上没有明确的标记。 将产生转基因小鼠，其中记忆B和T细胞将被 基因标记，因此可以明确地识别和检查。 免疫分化的研究得益于免疫调节剂的可用性。 克隆细胞系和嵌合分析方法。 扩展这些 有利于神经元分化的研究，方法将被开发 利用癌基因制备分化神经元的克隆衍生物， 一个精确的标记程序。 同时也是为了研究其他致命的 基因在神经元发育，嵌合分析的方法将是 开发 它将特别适用于分析的作用， 神经系统信号传导中的NF-κ B转录因子。