ROLE OF RNAI IN ADULT MAMMALIAN BRAIN

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。我们先前预测RNA干扰（RNAi）可能在成人大脑中生理性地发生，并参与调节与学习和记忆有关的基因表达变化。自这一预测以来，大量的microRNA已被证明存在于成人大脑中，与fmr蛋白相互作用并可能调节蛋白质合成。我们想了解Dicer、EIF-2c和其他参与RNA干扰的蛋白质是否在树突棘上表达，以及它们是否调节特定微小RNA和mRNA的局部合成。在UIC，我们正在对这个问题进行生物化学和行为研究，并试图在体内敲除Dicer。然而，我们试图利用NCMIR在形态学上定位Dicer和EIF-2c蛋白的专业知识，特别是在神经元激活和学习过程中。我们还需要使用原位杂交以高分辨率定位单个microRNA、microRNA前体及其mRNA靶标。最后，我们想利用共振能量转移寻找Dicer与其他蛋白质和RNA结合的动态变化。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We have previously predicted that RNA interference (RNAi) may occur physiologically in the adult brain and participate in regulating gene expression changes involved in learning and memory. Since this prediction, a large number of microRNAs have been demonstrated to be present in adult brain, interact with fmr protein and might regulate protein synthesis. We want to learn if Dicer, EIF-2c and other proteins involved in RNAi are expressed at dendritic spines, and if they regulate local synthesis of specific microRNAs and mRNAs. At UIC we are carrying out biochemical and behavioral studies of this question, as well as trying to knock out Dicer in vivo. However, we seek to take advantage of the expertise of NCMIR in localizing Dicer and EIF-2c proteins morphologically, particularly during neuronal activation and learning. We also need to localize individual microRNAs, microRNA precursors and their mRNA targets at high resolution using in situ hybridization. Finally, we would like to look for dynamic changes in the association of Dicer with other proteins and RNAs using resonance energy transfer.