Enhancing the Intracellular Functioning of anti-HIV RNAs

在培养的细胞中有效抑制HIV-1感染。我们将扩展对这些结构的分析， 原代CD 34+细胞和T淋巴细胞作为具体目标1的一部分。在目标2中，进一步分析HIV-1 RNA运输的功能作用 将被探索。具体来说，我们将研究2 '0-甲基共价骨架修饰 HN-1 RNA的R区的RNA由细胞小核仁RNA引导。我们将设计一个小的核仁RNA， 在HIV TAR和R13 E元件中，蛋白质相互作用的特定位点的甲基化。最后，将开发一种体内SELEX方案。 选择新的目标，在HIV-1和细胞RNA使用随机结合臂库的核仁定位核酶。的 在这一目标下提出的工作结果将扩展我们对HIV RNA和蛋白质通过Tht运输的功能作用的认识。 核仁，并提供新的核酶和其他抑制RNA用于抑制HIV-1感染。这项工作的总体目标是tc 通过使用核酶增强我们对HIV RNA定位的理解，并确定最佳的核酶-HIV靶向策略， 用于人类基因治疗。 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