Genetics

遗传学

• 批准号: 7663285
• 负责人: YOSEF REFAELI
• 金额: $ 15.36万
• 依托单位: NATIONAL JEWISH HEALTH
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7663285
• 关键词: AN-1; Antigen Receptors; Apoptosis Regulator; B-Lymphocytes; Cell Line; Cells; Development; Emerging Technologies; Family; Gene Expression; Generations; Genes; Genetic; Goals; Hematopoietic; Hematopoietic stem cells; Homeostasis; Lymphocyte; Lymphoid Cell; Mediating; Molecular; Mouse Strains; Mus; NF-kappa B; Nature; PTEN gene; Pathway interactions; Process; Production; Protein Family; Protein Overexpression; Receptors, Antigen, B-Cell; Research Personnel; Resources; Role; Signal Transduction; Specificity; Technology; Training; Training Support; Transgenic Mice; Viral; Viral Vector; autoreactive B cell; base; cDNA Expression; design; gain of function; genetic manipulation; improved; interest; killings; loss of function; member; new technology; pluripotency; programs; small hairpin RNA; tool; vector

This core aims to provide access, support and training for three new and emerging technologies for the projects described in this program. In addition, the core seeks to continue to develop and improve of the application of the three technologies described for the specific needs of the projects described in this program. The specific technologies include: a) the development of a comprehensive and technically advanced resource for the design, generation and production of viral-based shRNA vectors as well as retroviral approaches to manipulate ectopic gene expression in primary cells and cell lines of interest, b) the use of genetically modified mice for the efficient immortalization of autoreactive B-cells, and c) the use of novel technologies to conditionally immortalize long term hematopoietic stem cells in order to generate cell lines from transgenic mice of interest for these projects. The three specific aims of this core are: To develop a suite of viral vectors that will be useful for genetic manipulation of primary lymphoid cells. The use of viral mediated delivery of shRNA will be the primary tool for loss-of-function studies whereas the use of retroviral driven cDNA expression will be the primary means of gain-of-function studies in mice. 1. To use mice that overexpress MYC in an inducible, and B-cell specific manner to immortalize autoreactive B-cells (AN-1 cells). This approach will be important for Dr. Cambier's project and their attempts to determine the nature and specificity of the B-cell antigen receptors on normal AN1/T3 cells. 2. To define the hematopoietic pluripotency of conditionally immortalized long-term hematopoietic stem cell clones. The genetic pathways we will focus on will be of broad interest and application to the other projects in this program, and we will train and assist those investigators in this process as needed. This core will be important in allowing the different projects described in this program to define the molecular basis by which antigen receptor derived signals are able to regulate lymphocyte tolerance, homeostasis and survival. Depending on the specific circumstances, the antigen receptor can anergize, tolerize or kill lymphoid cells with the ultimate goal of generating an innocuous but useful repertoire. The projects described in this project will focus initially on the role of members of the Bcl-2 family in these functions of the antigen receptors.

这一核心旨在为以下三项新兴技术提供访问、支持和培训 本计划中描述的项目。此外，核心寻求继续发展和完善的 根据本文件中所述项目的具体需求，应用所述三项技术 程序。具体技术包括：a)开发一种全面的、技术上的 设计、生成和生产基于病毒的shRNA载体以及 逆转录病毒方法在原代细胞和感兴趣的细胞系中操纵异位基因表达，b) 利用转基因小鼠有效地使自身反应性B细胞永生化，以及c)使用 有条件地永生化长期造血干细胞以生成细胞的新技术 来自这些项目感兴趣的转基因小鼠的品系。该核心的三个具体目标是： 开发一套可用于初级淋巴细胞遗传操作的病毒载体。这个 使用病毒介导的shRNA传递将是功能丧失研究的主要工具，而使用 逆转录病毒驱动的c DNA表达将成为小鼠功能获得研究的主要手段。 1.使用高表达MYC的小鼠以可诱导的B细胞特异性方式永生化 自身反应性B细胞(AN-1细胞)。这种方法对Cambier博士的项目和他们的 试图确定正常AN1/T3上B细胞抗原受体的性质和特异性 细胞。 2.确定条件永生化的长期造血干细胞的造血多能性 细胞克隆。 我们将关注的遗传途径将对本研究的其他项目产生广泛的兴趣和应用。 计划，我们将根据需要培训和协助这些调查人员进行这一过程。这个核心将是 重要的是允许本计划中描述的不同项目定义分子基础 抗原受体衍生的信号能够调节淋巴细胞的耐受性、动态平衡和存活。 根据具体情况，抗原受体可以抵抗、耐受或杀死淋巴样细胞。 最终目标是产生一个无害但有用的曲目。本项目中描述的项目 首先将重点放在Bcl2家族成员在抗原受体的这些功能中的作用。