Genetics

该核心旨在为三项新兴技术提供获取、支持和培训， 本方案所述的项目。此外，核心寻求继续发展和改善的 本报告所述项目的具体需求所描述的三种技术的应用 程序.具体的技术包括：a）开发一个全面的，技术上 设计、生成和生产基于病毒的shRNA载体的先进资源，以及 逆转录病毒方法来操纵感兴趣的原代细胞和细胞系中的异位基因表达，B） 遗传修饰小鼠用于自身反应性B细胞的有效永生化的用途，和c）使用 有条件地使长期造血干细胞永生化以产生细胞的新技术 转基因小鼠的品系。这一核心的三个具体目标是： 开发一套可用于原代淋巴细胞遗传操作的病毒载体。的 使用病毒介导的shRNA递送将是功能丧失研究的主要工具， 逆转录病毒驱动的cDNA表达将是小鼠功能获得研究的主要手段。 1.使用以可诱导和B细胞特异性方式过表达MYC的小鼠来永生化 自身反应性B细胞（AN-1细胞）。这种方法对Cambier博士的项目及其 试图确定正常AN 1/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领域的其他项目产生广泛的兴趣和应用。 计划，我们将根据需要在这一过程中培训和协助这些调查人员。这个核心将是 重要的是允许在这个程序中描述的不同项目，以确定分子基础， 抗原受体衍生的信号能够调节淋巴细胞耐受性、体内平衡和存活。 根据具体情况，抗原受体可使淋巴细胞无反应、耐受或杀伤 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 将首先集中在Bcl-2家族成员在抗原受体的这些功能中的作用。

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.