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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要/摘要转录因子协调细胞中最动态的过程——从产生级联的抗病毒药物和抗生素抗性蛋白的分化和转移，即使是单一的丰度也会发生变化。转录因子可以诱导基因表达模式，从根本上改变转录组和细胞的生理学表明，转录因子对基因表达的影响是因子本身的丰度以及 DNA 中结合位点的丰度。 DNA结合位点增加，每个转录因子分子的作用减弱，然而，在细胞中，蛋白质。丰度变化而DNA结合位点的数量是静态的，因此转录因子的量是唯一的我们的目标是设计和测试一种能够产生丰富的分子技术。体内DNA中的转录因子结合位点，根据需要这些DNA分子会竞争占据。通过转录因子，从而拮抗它们对内源基因的作用。我们将设计逆转录元件，在诱导型和细胞类型的控制下逆转录目标位点这些元件可以作为合成的可编程部分整合到遗传电路中。生物学和对响应细胞状态的转录组进行控制的方法也成为可能。目前还没有小分子拮抗剂的转录因子的拮抗作用，我们将对此进行验证。方法在两个临床相关模型中进行：癌症进展和抗生素耐药性。