REGULATION OF RECEPTORS ON T LYMPHOCYTES

描述：森博士的长期目标是了解一个分子 使T细胞分化的连续步骤水平化。森博士 提出了确定调节转录的关键转录因子， 阶段特异性基因表达提供了一种重要机制， 看看分化过程。T细胞的调节 受体β链基因和CD 8 α链基因将被 作为双阴性和双阴性细胞中基因表达的探针， T细胞发育的阳性阶段。具体目标 本申请的目的是：1）研究生物学功能， J β 2-C β 2内含子中鉴定的启动子的调节。 有人提出，这种启动子可以调节TCR β重排， 通过在胚胎干细胞中缺失启动子来研究 细胞关键启动子因子将通过转染鉴定， 诱变和体外结合研究。 2)识别 在双阴性T细胞中激活TCR β增强子的因子 细胞一个核心增强子将被定义和分化阶段 将鉴定与这些序列相互作用的特定因子 通过比较突变小鼠和分期胎儿的提取物 胸腺细胞3)为了定义和表征CD 8 α基因调控 元素转基因小鼠将用于鉴定以下组合： 作为基因座控制的DNA酶I超敏位点 区域，并赋予发展适当的监管。的 结合遗传和生物化学实验， 对胸腺中的发育线索作出反应的因素， 从而为T细胞的操作提供靶点， 分化

DESCRIPTION: Dr. Sen's long term goal is to understand at a molecular level the sequential steps of T-cell differentiation. Dr. Sen proposes that identification of key transcription factors that regulate stage-specific gene expression provides an important mechanism to examine the differentiation process. The regulation of the T-cell receptor beta chain gene and the CD8 alpha chain gene will be studied as probes of gene expression in double negative and double positive stages of T-cell development, respectively. The Specific Aims of this application are: 1) To study the biological function and regulation of a promoter identified in the J beta 2-C beta 2 intron. It is proposed that this promoter may regulate TCR beta rearrangements, which will be investigated by deleting the promoter in embryonic stem cells. Critical promoter factors will be identified by transfection, mutagenesis and in vitro binding studies. 2) To identify the factors that activate the TCR beta enhancer in double negative T- cells. A core enhancer will be defined and differentiation stage specific factors that interact with these sequences will be identified by comparing extracts from mutant mice and staged fetal thymocytes. 3) To define and characterize CD8 alpha gene regulatory elements. Transgenic mice will be used to identify combinations of DNAase I hypersensitive sites that function as a locus control region and confer developmentally appropriate regulation. The combined genetic and biochemical experiments are expected to identify factors that respond to developmental cues in the thymus and, thereby, provide targets for the manipulation of T- cell differentiation.