Sensor-seq: A genome-wide biological measure of microRNA activity.

描述(申请人提供)：microRNA(MiRNA)是最近发现的一类有助于调节基因表达和控制细胞功能的调控RNA。已经发现了2000多个人类miRNAs。许多miRNAs具有肿瘤抑制或致癌功能，而miRNA调控失调在肿瘤的发生、转移甚至化疗耐药中起着关键作用。虽然人们已经对miRNA生物学有了很多了解，但随着新的miRNA基因的发现，对相关miRNA调控网络的注释变得越来越具有挑战性。一个关键的限制是缺乏高通量的生物学方法来对miRNA行为进行全基因组分析。在这份R33提案中，我们将开发和验证一种新的技术和方法，该技术和方法可用于：(1)在单个细胞分辨率下测量细胞内每个miRNA的活性， 以及(2)确定可用于改善针对自杀载体和正在开发的癌症治疗的溶瘤病毒的靶向的最佳合成miRNA结合位点。我们的技术将对扩大我们对癌症生物学的理解，产生研究甚至跟踪特定癌细胞亚群的工具，如癌症干细胞，筛选miRNA调节药物，以及开发更好地针对肿瘤细胞的新疗法具有重大实用价值。

DESCRIPTION (provided by applicant): microRNA (miRNA) are a recently uncovered class of regulatory RNAs which help to regulate gene expression, and control cell function. Over 2,000 human miRNAs have been discovered. Many miRNAs have tumor suppressor or oncogenic functions, and miRNA dysregulation has been shown to play a key role oncogenesis, metastasis, and even chemoresistance. Although much has been learned about miRNA biology, as the number of new miRNA genes have been discovered it has become increasingly challenging to annotate relevant miRNA regulatory networks. A critical limitation has been the lack of high-throughput, biological approaches for genome-wide analysis of miRNA behavior. In this R33 proposal, we will develop and validate a new technology and methodology that can be used to: (1) Measure the activity of each and every miRNA within a cell, at single cell resolution, and (2) Identify optimal synthetic miRNA binding sites that can be used to improve the targeting of suicide vectors and oncolytic viruses being developed for cancer therapies. Our technology will have major utility for expanding our understanding of cancer biology, for generating tools to study and even track specific cancer cell subsets, such as cancer stem cells, for screening for miRNA modulating drugs, and for developing novel therapies that better target tumor cells for destruction.