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.
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