Transcriptional Activation by Small RNA: Mechanism and Design Rules

描述（由申请人提供）：本申请的目的是建立一种由小RNA介导的基因调控的新范例。直到最近，小RNA一直被认为主要是作为一种进化上保守的基因沉默机制（称为RNAi）的介体发挥作用。我们和其他人发现，小双链RNA（dsRNA）靶向基因启动子序列可以在转录水平上以序列特异性方式诱导有效和延长的基因激活，我们将这种现象称为RNAa。尽管在活细胞中重编程基因表达具有巨大的潜力，但RNA的分子机制在很大程度上是未知的，靶点选择的规则仍然非常模糊。我们建议使用基因组学，生物信息学和分子生物学方法的组合来解决以下问题：如何实现启动子靶向dsRNA的转录激活？分子机器涉及什么？RNA是自然利用细胞的生理功能？控制dsRNA启动子靶向的灵敏度和特异性的一般规则是什么？了解RNA的分子机制并将其建立为一种新的范式，可能会改变基因功能研究和癌症等疾病治疗的方式。

DESCRIPTION (provided by applicant): The goal of this application is to establish a new paradigm of gene regulation mediated by small RNA. Until recently, small RNAs have been thought to function mainly as the mediators of an evolutionally conserved gene silencing mechanism known as RNAi. We and others discovered that small double stranded RNA (dsRNA) targeting gene promoter sequence can induce potent and prolonged gene activation at the transcriptional level and in a sequence-specific manner, a phenomenon we termed RNAa. Despite the enormous potential for reprogramming gene expression in living cells, the molecular mechanism of RNAa is largely unknown and the rules for target selection are still quite obscure. We propose to use a combination of genomic, bioinformatic, and molecular biology approaches to address the following questions: How is transcriptional activation by promoter-targeted dsRNA achieved? What are the molecular machines involved? Is RNAa naturally exploited by cells for physiological function? What are the general rules governing the sensitivity and specificity of dsRNA-promoter targeting? Understanding the molecular mechanism of RNAa and establishing it as a new paradigm are likely to transform the way gene function is studied and diseases such as cancer are treated.