Permanent Knockdown of MicroRNAs using a Lentiviral-based Anti-miR System

描述（由申请人提供）：一个由小rna介导的新的复杂基因调控层已经开始彻底改变我们对细胞如何在转录后水平调节基因表达的理解。大量的小调控rna被称为microrna (miRNAs)，其长度约为19-22个核苷酸，最近被发现在涉及多种生物过程的基因调控网络中发挥重要作用，包括干细胞分化、器官发育、信号传导、疾病和癌症。System Biosciences提出构建一种基于慢病毒的方法来表达反义microRNAs（“Anti-miRs”），该方法将在功能上灭活内源性microRNAs。这种类型的系统将使研究人员能够开始解开由microRNA控制的复杂调控回路的过程，从而更好地理解microRNA在细胞发育、细胞模式、信号传导和人类疾病中可能发挥的作用。在这个SBIR研究项目的第一阶段，System Biosciences将开发一个包含100个Anti-miR慢病毒表达构建体的小文库，并演示成功表达小反义microrna以及敲低靶向内源性microrna。公共卫生相关性：这项研究将为研究人员提供稳定和永久地敲除特定microRNA的方法，从而使公共卫生受益。这将使研究一个给定的microRNA在细胞发育、细胞模式、信号传导和人类疾病中可能发挥的所有作用成为可能。

DESCRIPTION (provided by applicant): A new and complicated layer of gene regulation mediated by small RNAs has begun to revolutionize our understanding of how the cell regulates gene expression on the posttranscriptional level. An abundant class of small regulatory RNAs, termed microRNAs (miRNAs), that are approximately 19-22 nucleotides in length have recently been identified to play major roles in gene regulatory networks involving multiple biological processes including stem cell differentiation, organ development, signaling, disease and cancer. System Biosciences proposes the construction of a lentiviral-based method for the expression of antisense microRNAs ("Anti-miRs") that will functionally inactivate endogenous microRNAs. This type of system will enable researchers to begin the process of unraveling the complex regulatory circuits governed by microRNAs - enabling a better understanding of the roles that the microRNA may be playing in cellular development, cell patterning, signaling, and human disease. During Phase I of this SBIR research project, System Biosciences will develop a small library of 100 Anti-miR lentiviral expression constructs and demonstrate the successful expression of the small antisense microRNAs as well as knockdown of targeted, endogenous microRNAs. PUBLIC HEALTH RELEVANCE: This research will benefit public health by providing researchers the means to stably and permanently knockdown a given microRNA. This will enable the investigation of all the roles that a given microRNA may be playing in cellular development, cell patterning, signaling, and human disease.