Systems analysis of mouse gastrulation

项目摘要 原肠形成是胚胎发生过程中产生三个主要胚种的基本事件。 从多能的外胚层分离出来。要理解这一复杂的过程，必须使用基于系统的 阐明控制密切协调的事件的遗传调控网络的方法 原肠形成过程中发生的图案化、分化和形态发生。在此应用程序中，我们将 使用复杂的计算工具从头开始重建监管网络，以调查 小鼠原肠发育。在我们的初步研究中，我们已经产生了一个基因调控网络(互动组) 为小鼠上皮细胞(EpiSC)使用无偏见的反向工程方法。利用基因表达 小鼠原肠周围胚胎产生的信号，我们询问了EpiSC相互作用组 确定原肠形成的候选主调控子，以及顺式调控序列基序和同源序列 在原肠形成过程中可能活跃的转录因子。因此，根据这些初步情况， 发现，我们假设我们的系统方法可以识别关键生物学的新的主控调节器 在小鼠原肠发育过程中。 我们现在将对支配老鼠的遗传调控网络进行全面的分析 原肠形成通过三个相互联系的特定目标：(1)鉴定小鼠上皮细胞的主要调控因子 原肠形成的无偏系统分析利用小鼠的原肠周围胚胎的信号来 询问EpiSC相互作用组以确定候选主调控子，然后是功能性 验证研究；(2)通过以下方式调节小鼠原肠发育的转录网络的分析 顺式调控基序和同源转录的计算识别和实验验证 可能在原肠形成中起作用的因素；以及(3)调节网络的功能分析 通过测试所推断的调控网络和主调节子的生物功能来原肠形成 来自原肠形成有缺陷的小鼠突变体的签名，并通过执行功能丧失分析 在活体小鼠胚胎中的候选主要调节器。综上所述，这些研究将提供新颖的 对对胚胎正常发育至关重要的中心生物过程的机械论洞察，以及 它的紊乱会导致严重的先天性缺陷。

Project Summary Gastrulation is the fundamental event during embryogenesis that generates the three primary germ layers from the pluripotent epiblast. To understand this intricate process, it is essential to use systems-based approaches to elucidate the genetic regulatory network that controls the tightly coordinated events of patterning, differentiation, and morphogenesis that take place during gastrulation. In this application, we will use sophisticated computational tools for the de novo reconstruction of regulatory networks to investigate mouse gastrulation. In our preliminary studies, we have generated a genetic regulatory network (interactome) for mouse epiblast stem cells (EpiSC) using unbiased reverse-engineering approaches. Using gene expression signatures generated from peri-gastrulation mouse embryos, we have interrogated the EpiSC interactome to identify candidate master regulators of gastrulation, as well as cis-regulatory sequence motifs and cognate transcription factors that are likely to be active during gastrulation. Therefore, based on these preliminary findings, we hypothesize that our systems approach can identify novel master regulators of key biological processes during mouse gastrulation. We will now pursue a comprehensive analysis of the genetic regulatory network that governs mouse gastrulation through three linked specific aims: (1) Identification of master regulators of mouse epiblast during gastrulation by unbiased systems analyses using signatures from peri-gastrulation mouse embryos to interrogate the EpiSC interactome for the identification of candidate master regulators, followed by functional validation studies; (2) Analysis of the transcriptional network that regulates mouse gastrulation by computational identification and experimental validation of cis-regulatory motifs and cognate transcription factors that are likely to function in gastrulation; and (3) Functional analyses of the regulatory network for gastrulation by testing the inferred regulatory network and biological functions of master regulators using signatures from mouse mutants that are defective in gastrulation, and by performing loss-of-function analyses of candidate master regulators in mouse embryos in vivo. Taken together, these studies will provide novel mechanistic insights into a central biological process that is essential for proper embryonic development, and whose perturbation can lead to major congenital defects.