Genetic and epigenetic mechanisms of developmental gene regulation

发育基因调控的遗传和表观遗传机制

• 批准号: 10226868
• 负责人: Daniel J McKay
• 金额: $ 38.25万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10226868
• 关键词: Animal Model; Animals; Area; Biochemical; Cell Maintenance; Cells; Chromatin; DNA; DNA Sequence; Defect; Development; Developmental Biology; Developmental Gene; Disease; Drosophila genus; Drosophila melanogaster; Enhancers; Epigenetic Process; Etiology; Funding; Funding Mechanisms; Gatekeeping; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genetic Transcription; Genome; Genomics; Genotype; Grant; Health; Heritability; Histones; Human; Knowledge; Life; Modernization; Molecular; Outcome; Pathway interactions; Play; Post-Translational Protein Processing; Process; Property; Regulation; Regulator Genes; Regulatory Element; Research; Role; Scientist; System; Testing; Time; Tissues; cell fate specification; developmental disease; fly; genetic resource; improved; insight; mutant; prevent; programs; spatiotemporal; student mentoring; transcription factor

Project Summary/Abstract My lab is broadly focused on developmental gene regulation. We study the transcriptional mechanisms that control determination and maintenance of cell fates over time. In particular, we seek to understand how access to regulatory DNA is spatiotemporally controlled. Because many developmental disorders and diseases acquired later in life are a consequence of gene regulatory defects, a better understanding of the underlying mechanisms is crucial for preventing diseases and improving their outcomes. We combine genomic, genetic and biochemical approaches in Drosophila melanogaster. The fly system offers multiple strengths, including a powerful ability to manipulate gene activity with temporal and spatial precision, small genome size, which affords cheap genomics, and a deep knowledge of the relevant genetic pathways, nearly all of which are conserved in humans. Research during the term of this grant will explore multiple questions at the core of modern developmental biology and the field of epigenetic gene regulation. We will investigate how the transcriptional programs underlying tissue identity are deployed in the proper temporal sequence during development. We have uncovered a temporal cascade of transcription factors which we hypothesize control the sequential activation and inactivation of transcriptional enhancers over time. We have termed these transcription factors as “chromatin gatekeepers” due to their requirement for opening and closing access to DNA regulatory elements. The mechanisms by which enhancers are inactivated, or “decommissioned” over time are particularly understudied. One of our objectives is to decipher these mechanisms. We will also investigate how information about decisions made earlier in development is propagated over time. A key to unlocking this question is a unique genetic resource we recently generated that enables us to directly test the function of histones. Histones are subject to a diverse array of post-translational modifications (PTMs) that are thought to serve as carriers of epigenetic information to regulate many DNA-templated processes. However, evidence supporting a functional role of histone PTMs in animals is largely correlative due to the difficulty in creating mutant histone genotypes in animals. Drosophila is distinct amongst animal models in that the histone genes reside at a single locus in the genome. We can replace the endogenous histone genes with tailor-made versions, thereby providing us with the first opportunity to distinguish between regulatory information that is directly encoded in the DNA sequence, and information that is epigenetically propagated. We will employ this approach to interrogate the molecular role that histone PTMs play in enhancer regulation and in heritable control of gene expression. MIRA funding would unify our research topics into a single funding mechanism. This will enable me to spend more time at the bench mentoring students, collaborating with other scientists, and exploring new and unexpected areas of research. Thus, MIRA support would maximize our ability to contribute to a mechanistic understanding of gene regulation, and the in the longer term, to leverage this insight toward understanding of disease etiology and treatment.

项目总结/摘要 我的实验室主要研究发育基因调控。我们研究转录机制， 控制细胞命运随时间的确定和维持。特别是，我们试图了解如何访问 是时空控制的。因为许多发育障碍和疾病 是基因调控缺陷的结果，对潜在机制的更好理解 对于预防疾病和改善其结果至关重要。我们结合了联合收割机基因组，遗传和生化 在果蝇中的应用。飞行系统提供了多种优势，包括强大的能力， 以时间和空间精度操纵基因活性，小基因组大小，这提供了廉价的基因组学， 以及对相关遗传途径的深入了解，几乎所有这些遗传途径都在人类中保守。 在此期间的研究将探讨现代发展的核心多个问题 生物学和表观遗传基因调控领域。我们将研究转录程序如何 潜在的组织身份在发育过程中以适当的时间顺序展开。我们有 我们发现了一个转录因子的时间级联，我们假设这些转录因子控制着连续的激活， 以及转录增强子随时间的失活。我们将这些转录因子称为“染色质 由于它们需要打开和关闭对DNA调控元件的访问，因此称为“看门人”。的 增强子随时间失活或“退役”的机制尤其未被充分研究。 我们的目标之一就是破译这些机制。我们还将研究决策信息 在早期的发展是随着时间的推移传播。解开这个问题的一个关键是一种独特的基因 我们最近产生的资源，使我们能够直接测试组蛋白的功能。组蛋白受 各种各样的翻译后修饰（PTM）被认为是表观遗传的载体， 信息来调节许多DNA模板化过程。然而，有证据表明， 动物中的组蛋白PTM在很大程度上是相关的，这是由于在动物中难以产生突变的组蛋白基因型。 动物果蝇在动物模型中是不同的，因为组蛋白基因位于染色体中的单个位点。 基因组我们可以用定制的版本替换内源性组蛋白基因，从而为我们提供 第一次有机会区分直接编码在DNA序列中的调节信息， 和表观遗传学传播的信息。我们将采用这种方法来询问分子作用 组蛋白PTM在增强子调节和基因表达的遗传控制中发挥作用。MIRA融资将 将我们的研究课题统一到一个单一的资助机制中。这将使我有更多的时间坐在板凳上 指导学生，与其他科学家合作，探索新的和意想不到的研究领域。 因此，MIRA的支持将最大限度地提高我们对基因调控机制的理解， 从长远来看，利用这种见解来理解疾病的病因和治疗。