Enhancing the Intracellular Functioning of anti-HIV RNAs

对培养细胞中HIV-1感染的有效抑制。我们将把对这些结构的分析扩展到 原代CD34细胞和T淋巴细胞作为特定目标的一部分1.在目标2中进一步分析HIV-1RNA运输的功能作用 通过核仁将被探索。具体地说，我们将研究2‘0-甲基共价主链修饰的可能性 在HN-1的R区，RNA由细胞内的小核仁RNA引导。我们将设计一种小的核仁RNA来误导2‘0 HIV TAR和R13E元件中特定蛋白质相互作用部位的甲基化。最后，将开发一种体内SELEX方案 使用核仁定位核酶的随机结合臂文库，在HIV-1和细胞RNA中选择新的靶点。这个 在这一目标中提出的工作结果应该扩大我们对HIV RNA和蛋白质通过 核仁，并提供新的核酶和其他抑制RNA，以抑制HIV-1感染。这项工作的总体目标是TC 通过使用核酶加深我们对HIV RNA定位的理解，并确定最佳的核酶-HIV靶向策略 未来在人类基因治疗中的应用。 我

potent inhibition of HIV-1 infection in cultured cells. We will extend our analyses of these constructs to primary CD34+ cells and T-lymphocytes as part of specific aim 1. In aim 2 further analyses of the functional role of HIV-1 RNA trafficking through the nucleolus will be explored. Specifically, we will investigate the possibility that 2'0-methyl covalent backbone modifications in the R region of HN-1 RNA are guided by a cellular small nucleolar RNA. We will engineer a small nucleolar RNA to misdirect 2'0 methylations to specific sites of protein interaction in the HIV TAR and Rl3E elements. Finally, an in vivo SELEX scheme will be developec to select for new targets, both in HIV-1 and cellular RNAs using a randomized binding arm library of nucleolar localized ribozymes. The results of the work proposed in this aim should extend our knowledge of the functional role of HIV RNA and protein trafficking through tht nucleolus, and provide new ribozymes and other inhibitory RNAs for inhibition of HIV-1 infection. The overall objective of this work is tc enhance our understanding of HIV RNA localization via the use of ribozymes, and to identify the best ribozyme-HIV target strategies for future use in human gene therapy. I