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.

项目摘要/摘要 生物体中的每个细胞几乎都具有完全相同的基因组信息，但是体内细胞 发展为截然不同的细胞身份。例如，一个细胞如何成为肝细胞和 另一个细胞变成了皮肤细胞？该终端分化为不同的单元格类型是由于正确的 该细胞类型的转录调节。转录因子（TF）是结合的调节蛋白 对于DNA的特定序列，响应元件，以打开或关闭该细胞类型的基因。自始至终 开发，TFS导航细胞的染色质景观并结合响应元素， 整合物理和基因特异性提示，指导精确的基因调节网络。对 TF动作是对正确响应元素的认可，并随后对其适当的调节 靶基因，同时不影响相邻基因的调节。但是，TF如何认识 正确理解正确的响应元件和靶基因。 TF中的突变及其反应 元素导致各种各样的人类疾病，因此了解这些基本机制 转录因子保真度对于我们对人类健康的理解至关重要。我们的长期目标是 表征TF响应元件识别和靶基因识别的机制 Dinstinct细胞类型。秀丽隐杆线虫为研究TF监管提供了独特的机会 紧凑的基因组。秀丽隐杆线虫中的基因间距离平均为2 kbp，这使得更容易 由于基因组复杂性降低而导致的响应元件与靶基因的相关性，而可能 需要有效的机制来掩盖异常TF动作。在此提案中，我们使用了一个良好的研究 NHR-25，具有两个哺乳动物直系同源物，NR5A1/SF-1和NR5A2/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 基因组和遗传技术的组合，利用新的基因组编辑方法，染色质 结合测定和秀丽隐杆线虫的独特基因组特征。这些实验将提供有关 TF在培训本科科学家时如何识别其响应元素和靶基因 尖端，基因组技术。