Resolving MicroRNA Targets

描述(由申请人提供)：小RNA生物学的性质还没有被很好地理解，有必要开发更多的工具来询问所涉及的因素。这份提案描述了我们努力开发新的方法来满足多巴胺受体表达神经元的这一需要。我们的策略是创造慢病毒传感器，‘感应’小RNA的表达水平。我们正在使用这一策略来询问存在于UTRs中的预测的信使核糖核酸靶标，并调节大脑中microRNAs的活动。我们还描述了我们将如何以混合池文库的形式为细胞生成数千个单独的慢病毒传感器--这对于剖析哺乳动物的非编码RNA功能来说是一个重大的进步。 与公共健康相关：在我们每个细胞内，都潜伏着一种暗物质。它以小RNA的形式存在，这些小RNA几乎可以调节我们生活的方方面面。在我们的大脑中，这些被称为microRNA的小RNA可能会调节与我们的行为交叉的过程，包括冲动、冒险、压力反应以及对药物滥用和成瘾的脆弱性。然而，人们对此知之甚少，以及其中涉及的基因是什么。这项提案描述了我们努力开发新的方法来询问大脑中小RNA的生物学。

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.