INTRACELLULAR SMALL RNA INHIBITORS OF HIV1

描述(改编自摘要)：细胞内表达 基于RNA的HIV-1功能抑制剂是一个潜在的强大工具，通过 哪种基因疗法可以用来对抗病毒复制。可能的 抑制剂包括反义RNA、催化RNA(核酶)，旨在 切断病毒RNA，或高亲和力RNA配体(适配子)，旨在结合 并抑制必要的病毒大分子的功能。在预赛中 实验中，“诱饵”RNA意在作为竞争性抑制因子 必需的HIV-1RNA结合蛋白REV已被证明是有效的。这个 这类RNA抑制剂的潜在靶点并不局限于 然而，通常对RNA有很高的亲和力。试管技术的出现 进化(或SELEX)技术使选择一系列 大分子多表位的高亲和力RNA配体(适配子) 抗原。An多个适配子抑制因子的同时表达 一系列病毒功能原则上可以通过以下方式克服突变逃逸 病毒。在拟议的研究中，首席调查员谈到 使用这种方法需要解决的主要问题。这些是：(1) 识别其他可提供高效表达的表达策略 人不同亚细胞位置稳定适配子的合成 细胞；以及(2)将适当的表达策略与现有和 新设计的抗HIV-1 RNA允许细胞合成自己的病毒 体内的抑制剂。将检测抗HIV-1适配子的表达 在稳定的细胞系中抑制感染的能力。 有效的策略将被优化和组合表达 将测试小RNA抑制剂对病毒的协同抑制作用 感染。

DESCRIPTION (adapted from the Abstract): The intracellular expression of RNA-based inhibitors of HIV-1 functions is potentially a powerful tool by which gene therapy might be used against viral replication. Possible inhibitors include antisense RNAs, catalytic RNAs (ribozymes) designed to cleave viral RNA, or high-affinity RNA ligands (aptamers) designed to bind and inhibit the function of essential viral macromolecules. In preliminary experiments, "decoy" RNAs meant to serve as competitive inhibitors of the essential HIV-1 RNA binding protein Rev have proven to be effective. The potential targets for such RNA inhibitors are not limited to proteins that normally have a high affinity for RNA, however. The advent of in vitro evolution (or SELEX) technology makes possible the selection of a series of high-affinity RNA ligands (aptamers) to multiple epitopes on macromolecular antigens. Simultaneous expression of multiple aptamer inhibitors of an array of viral functions could, in principle, overcome mutational escape by the virus. In the proposed research, the Principal Investigator addresses major problems to be solved using such an approach. These are: (1) identification of additional expression strategies that give efficient synthesis of stable aptamers in diverse subcellular locations in human cells; and (2) matching appropriate expression strategies with existing and newly devised anti-HIV-1 RNAs to allow cells to synthesize their own viral inhibitors in vivo. Expression of anti-HIV-1 aptamers will be tested for the ability to suppress infection in stably transfected cell lines. Effective strategies will be optimized and combinatorial expression of the small RNA inhibitors will be tested for synergistic inhibition of viral infection.