ANTIVIRAL HAIRPIN RIBOZYMES

核糖核酸酶，或称核酶，是一种非常有吸引力的分子。 作为序列特异性抗病毒治疗药物的潜在用途。这 该项目直接解决了关于以下两个中心科学问题 核酶--首先，核酶的分子结构如何导致 催化活性？第二，工程核酶能否被有效利用？ 在哺乳动物细胞内？我们的实验利用了催化作用 多功能50个核苷酸发夹状核酶。到目前为止的进展包括 一种新型高效体外选育技术的开发与利用 系统，并使用更传统的方法，来确定结构 以及对核酶，特别是其底物的生化要求 识别机制。本项目的具体目标是：(1) 确定底物结合所需的所有基本结构元件， RNA折叠，发夹状核酶的催化功能。(2)绘制地图 核酶和核酶-底物复合体的三级结构。(3) 表征和优化该酶的活性、选择性和 核酶在细胞质提取液中的稳定性。(四)严谨示范 并通过工程发夹状核酶优化细胞内催化。(5) 探索将核酶定向到特定细胞的两种新策略 地点。该项目将导致技术上的发展 合理设计治疗性核酶治疗慢性粒细胞白血病的能力 基本上是任何当前或未来的病毒性疾病。这样做的结果 该项目将对在以下领域工作的人具有直接重要性 生物分子催化，RNA结构，分子识别， 发展疗法、传染病和基因疗法。

RNA enzymes, or ribozymes, are fascinating molecules that are of great potential utility as sequence-specific antiviral therapeutic agents. This project directly addresses the two central scientific questions regarding ribozymes-- First, how does the molecular structure of ribozymes result in catalytic activity? Second, can engineered ribozymes be used effectively within the mammalian cell? Our experiments utilize the catalytically versatile 50 nt hairpin ribozyme. Progress to date includes the development and exploitation of a novel and powerful in vitro selection system, and the use of more traditional methods, to determine structural and biochemical requirements for the ribozyme, especially its substrate recognition mechanism. The Specific Aims of this project are to: (1) Identify all essential structural elements required for substrate binding, RNA folding, and catalytic function of the hairpin ribozyme. (2) Map the tertiary structure of the ribozyme and ribozyme-substrate complex. (3) Characterize and optimize the enzymatic activity, selectivity and stability of ribozymes in cytoplasmic extracts. (4) Rigorously demonstrate and optimize intracellular catalysis by engineered hairpin ribozymes. (5) Explore two novel strategies for directing ribozymes to specific cellular locations. The project will result in development of the technical capacity to rationally design therapeutic ribozymes for the treatment of essentially any current or future viral disease. The results of this project will be of immediate importance to those working in the areas of biomolecular catalysis, RNA structure, molecular recognition, developmental therapeutics, infectious disease, and gene therapy.