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

描述（由申请人提供）：微小RNA（miRNA）是最近发现的一类调节RNA，其有助于调节基因表达并控制细胞功能。人类已经发现了超过2,000种miRNAs。许多miRNA具有肿瘤抑制或致癌功能，并且miRNA失调已被证明在肿瘤发生、转移甚至化学抗性中起关键作用。虽然已经了解了很多关于miRNA生物学的知识，但随着新的miRNA基因的发现，注释相关的miRNA调控网络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分析miRNA的全基因组行为。在这项R33提案中，我们将开发和验证一种新的技术和方法，可用于：（1）以单细胞分辨率测量细胞内每个miRNA的活性， 和（2）鉴定最佳的合成miRNA结合位点，所述最佳的合成miRNA结合位点可用于改善正在开发的用于癌症治疗的自杀载体和溶瘤病毒的靶向。我们的技术将在扩大我们对癌症生物学的理解，生成研究甚至跟踪特定癌细胞亚群（如癌症干细胞）的工具，筛选miRNA调节药物，以及开发更好地靶向肿瘤细胞进行破坏的新疗法方面具有重要作用。

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.