RNAi-Mediated Gene Suppression in Adult Mammalian CNS

描述：(申请人提供) 我们推测，在生理条件下，RNA干扰(RNAi)可能发生 在成年哺乳动物中枢神经系统的条件下，它可能会引起长期的 抑制特定基因。如果RNAi确实发生了，它可能参与了 适应性、可塑性神经元反应，如基础学习和药物 加法。有人提出了一个项目来测试RNAi在学习中的假定作用， 这可以概括如下：首先，让小鼠接受嗅觉 学习范式。第二，找出在学习过程中被抑制的基因， 尤其是那些实验上与学习机制相关的。第三，做好准备 这些已识别的一个或多个mRNAs(siRNAs或 正义-反义杂交体)，并将它们引入幼小鼠的大脑中 诱导RNAi。这些小鼠的学习会得到促进吗？第四，获得 RNAi是内源性机制的生化证据 在正常情况下抑制其中至少一部分基因的表达(S) 学习。在这笔R21拨款中，为了获得作为初步证据所需的证据 评估这一假设，我们的目标是： 1.测试是否可以在实验中引发基因抑制 哺乳动物体内脑内的序列特异性方式。 2.测试siRNA转移是否可以成为一种有效的局部生产方法 在活体内注射到哺乳动物脑内时的基因抑制。 这些研究将开创将siRNA转移作为一种潜在的 选择性下调神经基因的表达，将为 为测试RNAi在学习和记忆中的作用奠定基础。

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.