Modular design of synthetic transcriptional regulators

描述（由申请人提供）：细胞命运由发育和细胞分化期间以程序化方式表达的基因组决定。通常情况下，在糖尿病和癌症等多种疾病中，转录调节因子发生故障会产生基因表达的异常模式，而这些模式正是疾病的核心。调控蛋白如何找到它们的结合位点并调控它们的靶基因仍然是该领域的中心问题。 我们结合联合收割机化学和生物学的方法来研究转录调节因子的其他棘手的功能。我们利用序列特异性DNA结合化合物（聚酰胺）来靶向特定的DNA序列，并且它们可以容易地被修饰以承载丰富的功能模块阵列。在拟议的工作中，我们将阐明合作DNA结合的基础外齿和Ultrabithorax，两个高度保守的发育调节。我们将应用这种理解来开发精确定制的合成调节因子，这些调节因子与细胞类型特异性转录因子合作靶向基因。最后，我们将测试我们的人工转录因子在细胞和生物体中调节基因的能力。 我们工作的最终目标是产生能够以所需方式调节靶基因表达的小分子。作为设计转录因子，这些分子将在解剖控制细胞命运和疾病的转录网络方面具有巨大价值。它们还具有作为由异常转录调节引起的多种疾病的治疗剂的潜力。

DESCRIPTION (provided by applicant): Cell fate is determined by sets of genes that are expressed in a programmed manner during development and cellular differentiation. Often, in diseases as diverse as diabetes and cancer, malfunctioning transcriptional regulators produce aberrant patterns of gene expression that are at the heart of the ailment. How regulatory proteins find their binding sites and regulate their targeted genes remains a central question in the field. We combine chemical and biological approaches to study otherwise intractable features of transcriptional regulators. We utilize sequence-specific DNA binding compounds (polyamides) to target specific DNA sequences, and they can be readily modified to bear a rich array of functional modules. In the proposed work, we will elucidate the basis of cooperative DNA binding by Extradenticle and Ultrabithorax, two highly conserved developmental regulators. We will apply that understanding to develop precisely tailored synthetic regulators that target genes cooperatively with cell-type specific transcription factors. Finally, we will test the ability of our artificial transcription factors to regulate genes in cells and in living organisms. The ultimate goal of our work is to generate small molecules that can regulate the expression of targeted genes in a desired manner. As designer transcription factors, these molecules will have tremendous value in dissecting transcriptional networks that govern cell fate and disease. They also have potential as therapeutic agents for a variety of diseases that are caused by aberrant transcriptional regulation.