INTRACELLULAR SMALL RNA INHIBITORS OF HIV1

描述（改编自摘要）： 基于RNA的HIV-1功能抑制剂是一种潜在的强大工具， 哪种基因疗法可以用来对抗病毒复制 可能 抑制剂包括反义RNA、设计成 切割病毒RNA或设计用于结合的高亲和力RNA配体（适体） 并抑制必需病毒大分子的功能。 初步 实验中，“诱饵”RNA意味着作为竞争性抑制剂， 必需的HIV-1 RNA结合蛋白Rev已被证明是有效的。 的 这种RNA抑制剂的潜在靶点不限于 通常对RNA有很高的亲和力。 试管婴儿的出现 进化（或SELEX）技术使得选择一系列 针对大分子上多个表位的高亲和力RNA配体（适体） 抗原 多个核酸适体抑制剂的同时表达 原则上，一系列病毒功能可以克服突变逃逸， 病毒 在拟议的研究中，主要研究者解决了 要解决的主要问题是采用这种方法。 这些是：（1） 鉴定另外的表达策略， 在人体不同亚细胞位置合成稳定的适体 细胞;以及（2）将适当的表达策略与现有的 新设计的抗HIV-1 RNA允许细胞合成自己的病毒 体内抑制剂。 将检测抗HIV-1适体的表达， 抑制稳定转染细胞系中感染的能力。 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， 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.