RNAi-Mediated Gene Suppression in Adult Mammalian CNS

描述：（申请人提供） 我们假设RNA干扰（RNAi）可能发生在生理条件下， 成年哺乳动物中枢神经系统的条件，它可能会导致长期的 抑制特定基因。如果真的发生了RNAi，它可能与 适应性、可塑性的神经元反应，如基础学习和药物 的增订条文提出了一个程序来测试RNAi在学习中的假定作用， 其可以总结如下：首先，使小鼠经受嗅觉刺激， 学习范式第二，识别在学习过程中受到抑制的基因， 尤其是那些与学习机制相关的实验。三是编制 这些鉴定的mRNA（siRNA或siRNA）中的一种或多种的双链RNA 有义-反义杂交体），并将其引入未处理小鼠的大脑中， 引发RNAi。这些老鼠的学习会变得更容易吗？第四，获得 生物化学证据表明，RNAi是内源性机制， 抑制这些基因中的至少一些基因在正常发育过程中的表达， 学习在这个R21补助金，以获得所需的初步证据， 评估这一假设，我们的目标是： 1.测试基因抑制是否可以在实验中引起， 序列特异性的方式在哺乳动物脑内的体内。 2.测试siRNA转移是否可以成为产生局部 当注射到哺乳动物脑内时的基因抑制。 这些研究将开创使用siRNA转移作为一种潜在的手段， 选择性敲低神经基因的表达， 这是测试RNAi在学习和记忆中作用的基础。

DESCRIPTION: (provided by applicant) We hypothesize that RNA interference (RNAi) may occur under physiological conditions in the adult mammalian CNS, and that it may cause long-term suppression of specific genes. If RNAi does occur, it may be involved in adaptive, plastic neuronal responses such as underlie learning and drug addition. A program is proposed to test the putative role of RNAi in learning, which can be summarized as follows: First, subject mice to an olfactory learning paradigm. Second, identify genes that are suppressed during learning, particularly those linked experimentally to learning mechanisms. Third, prepare double-stranded RNAs for one or more of these identified mRNAs (siRNAs or sense-antisense hybrids) and introduce them into the brains of naive mice to elicit RNAi. Will learning be facilitated in these mice? Fourth, obtain biochemical evidence that RNAi is the endogenous mechanism responsible for suppressing the expression of a least some of these gene(s) during normal learning. In this R21 grant, to gain evidence needed as a preliminary to evaluate this hypothesis, we aim to: 1. Test whether gene suppression can be elicited experimentally in a sequence-specific manner within mammalian brain in vivo. 2. Test whether siRNA transfer can be an effective method for producing local gene suppression when injected within mammalian brain in vivo. These studies will pioneer the use of siRNA transfer as a potential means of selectivity knocking-down expression of neural genes, and will lay the foundation for testing the role of RNAi in learning and memory.