The genetic and epigenetic mechanisms of phenotypic innovationhttps://apps.era.nih.gov/gm/reportCheckList.do?applicationID=9798249

项目总结 这项研究计划的目标是剖析新的调节电路和新特性背后的基因，以 更好地理解形态和细胞创新的遗传基础。每一种形态 结构或特征起源于过去的某个时间点，并在不同的进化路径下进化。然而， 目前尚不清楚一种新的性状是如何起源的，以及基因和调控网络是如何在空间上协调 发展新的细胞类型、组织和器官。确定推动和治理的流程 形态和功能的多样性和复杂性是理解生物进化的重要一步 复杂的生活。然而，我们对这一过程的理解仍然有限。这项研究的长期目标是 计划是从功能上描述新细胞团和新形态的分子遗传学基础 表型。中心假设是，从新的监管网络中涌现出的进化创新 取决于转录因子和增强子的变化。由包括单细胞RNA在内的初步数据指导 测序和成熟的理论，拟议的研究将使用一个 综合方法。我们将确定：1)果蝇新细胞团的调控网络创新，2) 负责转录因子表达变化和下游表达网络的增强子 修饰，以及3)新性状的基因调控基础。我们进行了单细胞测序和RNA 睾丸荧光原位杂交(FISH)发现果蝇分化出一个新的细胞团 物种。结合ATAC测序数据，我们将使用建模和功能研究来研究基因 新的细胞团的基础。根据这个假设，新的转录因子增强子(TF)是 ，我们将研究重复出现的小说特质的原因，并检验小说 调节电路对于一种新的特性是必不可少的。我们假设新的增强子或顺式调控基序 转录因子对于染色质可及性的全基因组水平的改变是必不可少的。为了测试它，我们将识别增强器 以及亲缘关系密切的物种之间的基序变化，以深入了解增强子和TF结合亲和力协同 进化论。这项研究将为转录调控网络的进化和 它们对新的形态和细胞特征的贡献。总而言之，我们的综合方法将有助于 阐明新的调控回路的起源和进化及其对表型的贡献 创新。

PROJECT SUMMARY The goal of this research proposal is to dissect the novel regulatory circuits and genes underlying novel traits, to get a better understanding of the genetic basis of morphological and cellular innovation. Every morphological structure or trait originated at some time point in the past and evolved under various evolutionary paths. However, it is unknown how a novel trait originates and how gene and regulatory networks spatially orchestrate the development of the novel cell types, tissues, and organs. Identifying the processes driving and governing morphological and functional diversity and complexity is a major step towards understanding the evolution of complex life. However, our understanding of this process is still limited. The long-term goal of this research program is to functionally characterize the molecular genetic basis of novel cell clusters and novel morphological phenotypes. The central hypothesis is that evolutionary innovations emerging from novel regulatory networks depend on changes in transcription factors and enhancers. Guided by preliminary data including single cell RNA sequencing and well-established theories, the proposed research will test the central hypothesis using an integrative approach. We will determine: 1) regulatory network innovation in novel cell clusters in Drosophila, 2) enhancers responsible for transcription factor expression changes and downstream expression network modification, and 3) the genetic regulatory basis of a novel trait. We performed single-cell sequencing and RNA fluorescent in situ hybridization (FISH) on testis and found a novel cell cluster differentiated between Drosophila species. Combined with ATAC-sequencing data, we will use modeling and functional studies to study the genetic basis of the novel cell cluster. Following this hypothesis that novel enhancers of transcription factors (TFs) are essential for novel traits, we will study the cause of a recurrent novel trait and test the hypothesis that a novel regulatory circuit is essential for a novel trait. We hypothesize that novel enhancers or cis-regulatory motifs of TFs are essential for whole-genome level changes in chromatin accessibility. To test it, we will identify enhancer and motif changes between closely related species to provide insights into enhancer and TF binding affinity co- evolution. This study will provide important insights into the evolution of transcription regulatory networks and their contributions to novel morphological and cellular traits. Altogether, our integrative approach will help to elucidate the origination and evolution of novel regulatory circuits and their contributions to phenotypic innovation.