RetroDecoys: Temporally-regulated and cell type selective technology for transcriptional control

RetroDecoys：用于转录控制的时间调节和细胞类型选择性技术

• 批准号: 10373249
• 负责人: Seth Lawler Shipman
• 金额: $ 23.63万
• 依托单位: J. DAVID GLADSTONE INSTITUTES
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-15 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10373249
• 关键词: Address; Antibiotic Resistance; Antibiotics; Antiviral resistance; Automobile Driving; Bacteria; Bacteriophages; Binding; Binding Sites; Biological; Biological Assay; Biotechnology; Cancer Model; Cell Differentiation process; Cell physiology; Cells; Cellular Assay; Clinical; Complex; DNA; DNA Binding; DNA Sequence; DNA amplification; Elements; Engineered Gene; Engineering; Escherichia coli; Eukaryotic Cell; Future; Gene Expression; Gene Expression Profile; Genes; Genetic; Genetic Transcription; Genome; Genomics; Goals; Growth; Malignant Neoplasms; Molecular; Multi-Drug Resistance; Neoplasm Metastasis; Oligonucleotides; Oncogenic; Pathway interactions; Pharmaceutical Preparations; Physiology; Play; Process; Production; Prokaryotic Cells; Proteins; RNA-Directed DNA Polymerase; Regulation; Research; Retroelements; Role; STAT3 gene; Single-Stranded DNA; Site; Synthetic Genes; System; Technology; Testing; Therapeutic; Time; Tissue Engineering; Titrations; Transcription Coactivator; Transcriptional Regulation; Viral; Work; antagonist; cell type; cellular engineering; clinically relevant; efficacy validation; functional genomics; in vivo; mutant; novel; novel therapeutics; nuclease; programs; promoter; small molecule; synthetic biology; tool; transcription factor; transcriptome; tumor progression

Project Summary/Abstract Transcription factors orchestrate the most dynamic processes in cells – from producing cascades of antiviral and antibiotic resistance proteins to differentiation and metastasis. A change in the abundance of even a single transcription factor can induce a pattern of gene expression that fundamentally alters the transcriptome and physiology of a cell. Interestingly, the effect that a transcription factor has on gene expression is a function of both the abundance of the factor itself as well as the abundance of its binding site in DNA. As the number of DNA binding sites increases, the effect of each transcription factor molecule decreases. Yet, in cells, protein abundance changes and the number of DNA binding sites is static, so the transcription factor amount is the only point of dynamic control. We aim to engineer and test a molecular technology that will produce abundant transcription factor binding sites in DNA in vivo, on demand. These DNA molecules will compete for occupancy by the transcription factors, thereby antagonizing their action on endogenous genes. To build this technology, we will engineer retroelements to reverse transcribe the target site under the control of inducible and cell-type specific promoters. These elements can be integrated into genetic circuits as programmable parts for synthetic biology and a means to exert control over a transcriptome that is responsive to a cell's state. They also enable antagonism of transcription factors that currently have no small molecule antagonist. We will validate this approach in two clinically relevant models: cancer progression and antibiotic resistance.

项目总结/摘要 转录因子协调细胞中最动态的过程-从产生抗病毒和抗病毒的级联反应， 抗生素抗性蛋白的分化和转移。即使是一个单一的 转录因子可以诱导基因表达的模式，从根本上改变转录组， 细胞的生理学。有趣的是，转录因子对基因表达的影响是一个功能， 因子本身的丰度以及其在DNA中结合位点的丰度。的数量 DNA结合位点增加，每个转录因子分子的作用降低。然而，在细胞中， 丰度的变化和DNA结合位点的数量是静态的，所以转录因子的数量是唯一的 动态控制点。我们的目标是设计和测试一种分子技术， 根据需要，在体内DNA中的转录因子结合位点。这些DNA分子会为了占据 通过转录因子，从而拮抗它们对内源基因的作用。为了建立这项技术， 我们将设计逆转录元件，在诱导型和细胞型的控制下逆转录靶位点， 特异性启动子这些元件可以集成到遗传电路中，作为合成的可编程部件。 生物学和一种手段来施加控制转录组是响应细胞的状态。它们还支持 目前没有小分子拮抗剂的转录因子的拮抗作用。我们会证实这一点 在两个临床相关的模型：癌症进展和抗生素耐药性的方法。