Resolving MicroRNA Targets

描述（由申请人提供）：小RNA生物学的性质尚不清楚，需要开发额外的工具来询问所涉及的因素。该提案描述了我们为实现多巴胺受体表达神经元的这一需求而开发新方法的努力。我们的策略是创造慢病毒传感器来“感知”小rna的表达水平。我们正在使用这种策略来询问utr中存在的预测mRNA目标，并调节大脑中microrna的活性。我们还描述了我们将如何以混合库的形式生成数千个单独的慢病毒传感器到细胞-这是解剖哺乳动物非编码RNA功能的重大进步。

DESCRIPTION (provided by applicant): The nature of small RNA biology is not well understood, and there is a need to develop additional tools to interrogate the factors involved. This proposal describes our efforts to develop novel methods for accomplishing this need in dopamine receptor expressing neurons. Our strategy is to create lentiviral sensors that 'sense' the expression level of small RNAs. We are using this strategy to interrogate the predicted mRNA targets present within UTRs, and also to modulate the activity of microRNAs in the brain. We also describe how we will generate thousands of individual lentivirus sensors to cells, in the form of a mixed-pool library- a significant advancement for the dissection of noncoding RNA function in mammals. PUBLIC HEALTH RELEVANCE: Inside each of our cells, there is a dark matter lurking. It takes the form of small RNAs, which have the power to regulate almost every aspect of our lives. Inside our brains, these small RNAs- known as microRNAs- likely regulate processes that intersect with our behaviors, including impulsivity, risk taking, stress responsivity, and vulnerability to drug abuse and addiction. However, little is known about how this works, and what are the genes involved. This proposal describes our efforts to develop novel methods for interrogating the biology of small RNAs in the brain.