Synthetic modulators of transcription

转录的合成调节剂

• 批准号: 6924713
• 负责人: MOTONARI UESUGI
• 金额: $ 24.83万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-01 至 2008-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6924713
• 关键词: DNA binding protein; bioengineering /biomedical engineering; biomimetics; biotechnology; enzyme linked immunosorbent assay; fluorescence microscopy; gene induction /repression; genetic manipulation; genetic transcription; human tissue; laboratory mouse; microarray technology; mitogens; nuclear magnetic resonance spectroscopy; nucleic acid sequence; organic chemicals; polymerase chain reaction; protein protein interaction; reporter genes; small molecule; transcription factor; vascular endothelial growth factors; western blottings

DESCRIPTION (provided by applicant): Regulation of gene expression touches almost everything in eukaryotic biology and often generates drastic phenotypes in living organisms. Precise exogenous control of transcription by cell-permeable synthetic molecules represents a challenge in medicinal and bioorganic chemistry. Here we propose to develop cell-permeable organic compounds that mimic transcription factors. Naturally occurring transcription factors typically have two separable domains for binding specific DNA sequences and for activating transcription through protein-protein interactions. We plan to create artificial transcription factors by replacing these two polypeptide domains with synthetic molecules. As an activation module, a wrench-shaped molecule we named "wrenchnolol" will be used, while a Dervan's hairpin polyamide molecule will be used as a DNA-binding module. The two cell-permeable modules will be coupled with each other at different positions and with different linkers, and the synthesized molecules will be assayed for their ability to activate transcription of a reporter gene in vitro, in cells, and in mice. We will also attempt to activate endogenous genes by synthetic transcription factors. We plan two approaches to achieving this challenging goal. In one, we will design and synthesize a synthetic molecule that mimics an endogenous transcription factor. As a proof of the concept, we plan to design a mimetic of MyoD, a transcription factor that induces myogenesis. In the other approach, a set of ten synthetic transcription factors will be designed in such a way that they synergistically activate transcription of a single gene. As a proof of the concept, we plan to activate the gene of VEGF-A, an endothelial cell-specific mitogen that is a key inducer of new blood vessel growth.

描述（由申请人提供）：基因表达的调节几乎触及真核生物学中的所有事物，并且经常在生物体中产生剧烈的表型。细胞可渗透的合成分子对转录的精确控制代表了药物和生物有机化学的挑战。在这里，我们建议开发可模拟转录因子的可渗透性有机化合物。天然发生的转录因子通常具有两个可分离域，用于结合特定的DNA序列，并通过蛋白质 - 蛋白质相互作用激活转录。我们计划通过用合成分子代替这两个多肽结构域来创建人工转录因子。作为激活模块，将使用我们称为“扳手”的扳手形分子，而Dervan的发夹聚酰胺分子将用作DNA结合模块。这两个可渗透的模块将在不同位置和不同的接头上相互耦合，并且将分析合成分子，以在细胞和小鼠中激活细胞，细胞中的报告基因转录的能力。我们还将尝试通过合成转录因子激活内源基因。我们计划实现这一具有挑战性的目标的两种方法。在一个中，我们将设计和合成一个模拟内源转录因子的合成分子。作为概念的证明，我们计划设计一种模仿Myod，这是诱导肌发生的转录因子。在另一种方法中，将设计一组十个合成转录因子，以使它们协同激活单个基因的转录。作为概念的证明，我们计划激活VEGF-A的基因，VEGF-A的基因是一种内皮细胞特异性有丝分裂原，是新血管生长的关键诱导剂。