Spatial Organization of Gene Expressoin

描述(由申请人提供)：这个项目的长期目标是对活细胞中RNA聚合酶II转录和前信使核糖核酸加工的调控所涉及的空间和时间事件有一个完整的了解。在活细胞和生物体的背景下理解这些参数将为确定与各种疾病状态相关的细胞或组织中发生的这些事件的变化提供基础。短期目标将利用先前开发的活细胞基因表达系统来解决活细胞中特定遗传位点在G2/M和M/G1转变时是如何调节的。本课程将研究基因表达相关因素的时空动态，以及在有丝分裂过程中基因转录状态如何传递到子代细胞。此外，该系统将用于研究与mRNP出口机制的部件何时何地与新生的mRNPs相关的关键悬而未决的问题，以及mRNPs如何通过人类细胞的核孔复合体运输。最后，将启动一系列高度创新的实验，将实时基因表达研究带入活动物的背景中，以研究皮肤分化时组织特异性动力学和基因表达的变化。这种方法将提供对组织内基因表达的重要洞察，并允许人们直接访问在体内分化和形态发生的背景下的核动力学。总之，这些研究将为我们理解与基因表达有关的核的功能组织提供重要的见解。核组织中的异常有可能导致基因表达的改变，其最终产物可能是与人类疾病相关的许多病理之一。这些研究将使我们更接近通过体内分子成像和动力学来研究和评估疾病模型的重要一步。

DESCRIPTION (provided by applicant): The long-term goal of this project is to develop a complete understanding of the spatial and temporal events involved in the regulation of RNA polymerase II transcription and pre-mRNA processing in living cells. An understanding of these parameters in the context of the living cell and organism will provide the basis to identify changes in these events that occur in cells or tissues associated with various disease states. The short-term goals will utilize a previously developed live cell gene expression system to address how a specific genetic locus is modulated at the G2/M and M/G1 transitions in living cells. The spatial and temporal dynamics of factors involved in gene expression will be examined as well as how the transcriptional status of a gene is transmitted to daughter cells during mitosis. In addition, this system will be used to study critical unanswered questions relating to where and when components of the mRNP export machinery associate with nascent mRNPs and how mRNPs are transported through the nuclear pore complex in human cells. Finally, a series of highly innovative experiments will be initiated to bring real-time gene expression studies into the context of a living animal in order to study tissue specific dynamics and changes in gene expression upon skin differentiation. This approach will provide significant insight into gene expression within a tissue and allow one to directly access nuclear dynamics in the context of differentiation and morphogenesis in vivo. Together, these studies will provide significant insight into our understanding of the functional organization of the nucleus with regard to gene expression. Aberrations in nuclear organization have the potential to result in alterations in gene expression, the end product of which may be one of the many pathologies associated with human disease. These studies will bring us a major step closer to studying and evaluating disease models by in vivo molecular imaging and dynamics.