Biological Validation Development

项目摘要/摘要-生物验证开发 本模块的最终目标是演示我们能够预测染色体位置 以及来自全基因组、DamID和TSA-Seq作图的相对于不同核室的轨迹 数据，并确定此染色体定位的生物学意义。 我们将通过三大目标实现这些目标。每一次都将涉及以下迭代周期 实验和计算机建模。第一个是验证对染色体定位的预测和 来自全基因组DAMID和TSA-SEQ数据的关于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，以便我们可以估计接触频率和 分别考虑相对于每个主要核舱室的距离分布。更多 雄心勃勃地，这些校准数据将被用作DAM模块中的计算机建模的输入：测绘数据 因为多个核隔室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。这些预测随后将通过直接显微镜进行验证。 并用于改进我们的预测性建模。 第二，我们的目标是识别负责靶向的dna区域，并最终识别顺式元件。 从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 不同核室附近染色体定位的功能后果。我们会这么做的 通过一种作为染色体位置函数的功能分析的活细胞读数的新组合 通过刻意地重新连接内源染色体的轨迹，然后读出 使用分子方法。 更具体地说，我们将重点关注以下具体目标：1：验证核内染色体 位置取决于DamID和TSA区段基因组图谱数据；2：测试预测 建立核内染色体区划的机制；3：测试预测 染色体定位对不同核区的功能影响 这些目标的实现应使我们有能力和信心解释 我们观察到的基因组组织相对于核隔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的作用，但 在建立和维持组织特有的基因表达模式方面，以前没有意识到的作用。 因此，本模块的工作将对我们新地图技术的未来应用至关重要 有助于更好地了解细胞在正常发育和人类疾病中的功能。

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.