Genomic Analysis into Transcriptional Regulation of Cell Identity

细胞身份转录调控的基因组分析

• 批准号: 10513735
• 负责人: Deborah Thurtle-Schmidt
• 金额: $ 38.05万
• 依托单位: DAVIDSON COLLEGE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10513735
• 关键词: Affect; Amino Acid Sequence; Amino Acids; Binding; Binding Proteins; Binding Sites; Biological Assay; C. elegans genome; Caenorhabditis elegans; Cell Lineage; Cell physiology; Cells; Chromatin; Clinical; Cues; DNA; DNA Binding; DNA Binding Domain; DNA Sequence; DNA-Binding Proteins; Development; Drosophila genus; Epithelial Cells; Gene Expression; Gene Expression Regulation; Gene Targeting; Genes; Genetic; Genetic Screening; Genetic Techniques; Genetic Transcription; Genome; Genomics; Goals; Health; Hepatocyte; Human; Human Genome; In Vitro; Ligand Binding; Maintenance; Methods; Modeling; Molecular; Mutation; NR5A2 gene; Normal Cell; Nuclear Hormone Receptors; Organism; Orthologous Gene; Phenotype; Physiological; Point Mutation; Post-Translational Protein Processing; Prevention; Proteins; RNA Interference; Regulation; Response Elements; Role; SF1; Scientist; Signal Transduction; Site; Skin; Techniques; Testing; Training; Transcriptional Regulation; United States National Institutes of Health; Variant; cell type; college; design; disorder prevention; embryo cell; experimental study; gene regulatory network; genetic corepressor; genetic regulatory protein; genome editing; human disease; innovation; insight; mutant; novel; promoter; recruit; screening; transcription factor; undergraduate student

Project Summary/Abstract Every single cell in an organism has almost the exact same genomic information, yet cells in the body develop into vastly different cell identities. For example, how does one cell become a liver cell and another cell become a skin cell? This terminal differentiation into distinct cell types is due to correct transcriptional regulation for that cell type. Transcription factors (TFs) are regulatory proteins that bind to specific sequences of DNA, response elements, to turn on or off genes for that cell type. Throughout development, TFs navigate the chromatin landscape of the cell and bind response elements, integrating physiological and gene-specific cues, directing precise gene regulatory networks. Critical to TF action is recognition of the correct response element and subsequent regulation to its appropriate target gene, while not affecting the neighboring genes’ regulation. However, how a TF recognizes the correct response element and target gene is not well understood. Mutations in TFs and their response elements result in a wide array of human diseases, thus understanding these basic mechanisms of transcription factor fidelity is critical to our understanding of human health. Our long-term goal is to characterize the mechanism of TF response element recognition and target gene recognition in dinstinct cell types. C. elegans provides a unique opportunity to study TF regulation due to its compact genome. Intergenic distance in C. elegans is on average 2 Kbp, which allows for easier correlation of response element to target gene due to the reduced genomic complexity while likely requiring efficient mechanism to shield aberrant TF action. In this proposal we use a well-studied nuclear hormone receptor, NHR-25, with two mammalian orthologs, NR5A1/SF-1 and NR5A2/SF-2, to interrogate TF binding and cell-type specific target gene regulation. We will do so through a combination of genomic and genetic techniques, exploiting new genome editing methods, chromatin binding assays, and unique genomic features of C. elegans. These experiments will provide insight into how TFs recognize their response element and target gene while training undergraduate scientists in cutting-edge, genomic techniques.

项目总结/摘要 生物体中的每一个细胞都有几乎完全相同的基因组信息， 发展成截然不同的细胞身份例如，一个细胞如何变成肝细胞， 另一个细胞变成皮肤细胞这种终末分化为不同的细胞类型是由于正确的 这种细胞类型的转录调控。转录因子（TF）是一种调节蛋白， 到特定的DNA序列，反应元件，来打开或关闭该细胞类型的基因。在整个 在发育过程中，TF导航细胞的染色质景观并结合反应元件， 整合生理和基因特异性线索，指导精确的基因调控网络。的关键 TF动作是识别正确的反应元件并随后对其进行适当的调节 目的基因，而不影响相邻基因的调控。然而，TF如何识别 正确的反应元件和靶基因还没有很好地理解。TF中的突变及其反应 元素导致各种人类疾病，从而了解这些基本机制， 转录因子的保真度对我们理解人类健康至关重要。我们的长期目标是 研究TF反应元件识别和靶基因识别的机制， 不同的细胞类型。C.线虫提供了一个独特的机会，研究TF调节，由于其 紧凑的基因组C. elegans平均为2 Kbp，这使得更容易 由于降低的基因组复杂性，响应元件与靶基因的相关性可能 需要有效的机制来屏蔽异常的TF作用。在这个建议中，我们使用一个经过充分研究的 核激素受体NHR-25与两个哺乳动物直系同源物NR 5A 1/SF-1和NR 5A 2/SF-2， 询问TF结合和细胞类型特异性靶基因调节。我们将通过一个 基因组和遗传技术的结合，开发新的基因组编辑方法，染色质 结合试验和C.优雅的这些实验将提供洞察力， TF如何识别它们的反应元件和靶基因，同时培训本科科学家， 尖端的基因组技术