Biological Validation Development

项目总结/摘要-生物学确认开发 本模块的最终目标是证明我们能够预测染色体位置 和相对于来自全基因组、DamID和TSA-Seq作图的不同核隔室的轨迹 数据并确定该染色体定位的生物学意义。 我们将通过三个主要目标来实现这些目标。每一个都将涉及迭代循环， 实验和计算机建模。首先是验证染色体定位的预测， 从全基因组DamID和TSA-Seq数据的主要核隔室的动力学。这 包括最初校准我们基因组图谱的输出，这样我们就可以估计接触频率， 距离分布相对于每一个主要的核隔间单独考虑。更 雄心勃勃的是，这些校准数据将用作DAM模块中计算机建模的输入：映射数据 多核区室的基因组数据将与其他基因组数据相结合，以产生更准确的预测。 染色体的位置和轨迹。然后这些预测将通过直接显微镜进行测试 观察并用于改进我们的预测模型。 其次，我们的目标是确定DNA区域，并最终顺式元件，负责靶向 染色体区域到特定的核隔室。我们的第三个也是最后一个目标是预测和测试 不同核区室附近染色体位点定位的功能后果。我们会这样做 通过作为染色体位置的函数的功能测定的活细胞读出的新组合 并通过故意重新布线内源染色体的轨迹，然后读出 使用分子方法。 更具体地说，我们将集中在以下具体目的：1：染色体的核内染色体 位置作为DamID和TSA区室全基因组作图数据的函数; 2：测试预测 建立核内染色体区室化的机制; 3：试验预测 染色体定位于不同核区室的功能后果 这些目标的完成应该使我们有能力和信心来解释 基因组组织相对于我们观察到的核区室的变化的生物学意义， 不同的细胞和组织类型。我们预计，这一级别的核组织将发挥关键作用， 在建立和维持基因表达的组织特异性模式中以前未实现的作用。 因此，本单元的工作对于我们新的测绘技术的未来应用至关重要 对正常发育和人类疾病中细胞功能的理解。

PROJECT SUMMARY / ABSTRACT – BIOLOGICAL VALIDATION DEVELOPMENT The ultimate goals of this Module are to demonstrate that we are able to predict chromosome locations and trajectories relative to different nuclear compartments from genome-wide, DamID and TSA-Seq mapping data AND to identify the biological significance of this chromosome positioning. We will accomplish these goals through three major aims. Each will involve iterative cycles of experimentation and computer modeling. The first is to validate predictions for chromosome positioning and dynamics with respect to major nuclear compartments from genome-wide DamID and TSA-Seq data. This involves initially calibrating the output of our genome maps, such that we can estimate contact frequencies and distance distributions relative to each of the major nuclear compartments considered separately. More ambitiously, this calibration data will be used as input for computer modeling in the DAM Module: mapping data for multiple nuclear compartments will be combined with other genomic data to yield more accurate predictions of chromosome location and trajectories. These predictions will then be tested by direct microscopy observations and used to refine our predictive modeling. Second, we aim to identify DNA regions, and ultimately cis elements, responsible for targeting chromosome regions to specific nuclear compartments. Our third and final goal is to predict and test the functional consequences of chromosome loci localization near different nuclear compartments. We will do this through a novel combination of live-cell readout of functional assays as a function of chromosome position AND through a deliberate, rewiring of the trajectories of endogenous chromosomes, followed by readouts using molecular methods. More specifically, we will focus on the following Specific Aims: 1: Validate intranuclear chromosome position as a function of DamID and TSA compartment genome-wide mapping data; 2: Test predicted mechanisms by which intranuclear chromosome compartmentalization is established; 3: Test predicted functional consequences of chromosome localization to different nuclear compartments Completion of these Aims should provide us with the capability and confidence for interpreting the biological significance of changes in genome organization relative to nuclear compartments that we observe in different cell and tissue types. We anticipate that this level of nuclear organization plays a critical but previously unrealized role in establishing and maintaining tissue-specific patterns of gene expression. Therefore the work in this Module will be critical for the future application of our new mapping technologies towards the improved understanding of cell function in normal development and human disease.