Spatial Organization of Gene Expressoin

描述（由申请人提供）：该项目的长期目标是全面了解活细胞中RNA聚合酶II转录和前mRNA加工调节所涉及的空间和时间事件。在活细胞和生物体的背景下对这些参数的理解将为识别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特定遗传位点如何在活细胞中的G2/M和M/G1转换中进行调节。参与基因表达的因素的空间和时间动态将被检查，以及如何在有丝分裂过程中基因的转录状态被传递到子细胞。此外，该系统还将用于研究关键的未回答的问题，这些问题涉及mRNP输出机制的组成部分在何处以及何时与新生mRNP相关联，以及mRNP如何通过人类细胞中的核孔复合物进行运输。最后，将启动一系列高度创新的实验，将实时基因表达研究纳入活体动物的背景下，以研究皮肤分化后的组织特异性动力学和基因表达变化。这种方法将提供对组织内基因表达的重要见解，并允许人们在体内分化和形态发生的背景下直接访问核动力学。总之，这些研究将为我们理解基因表达方面的细胞核功能组织提供重要的见解。核组织的畸变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。这些研究将使我们在通过体内分子成像和动力学研究和评估疾病模型方面迈出重要一步。

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.