DESIGN OF OPTIMAL RIBOZYMES FOR CLEAVAGE OF HIV RNA

用于切割 HIV RNA 的最佳核酶的设计

• 批准号: 2065524
• 负责人: OLKE C UHLENBECK
• 金额: $ 15.76万
• 依托单位: UNIVERSITY OF COLORADO
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-03-01 至 1998-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2065524
• 关键词: RNA binding protein; enzyme activity; enzyme inhibitors; enzyme mechanism; genetic library; human immunodeficiency virus; molecular cloning; neomycin; nucleic acid chemical synthesis; nucleic acid sequence; polymerase chain reaction; protein engineering; ribozymes; virus RNA

The goal of this project is to develop the use of RNA enzymes as reagents for the site specific cleavage (a consequent inactivation) of HIV and other high molecular weight RNAs in vitro and in vivo. Recent success in several labs using the hammerhead ribozyme to inactivate a number of RNAs (including HIV sequences) in bacterial, plant and animal cells emphasize the value of this emerging technology. This project will focus on refining and expanding the biochemical description of the cleavage of small and large RNAs by the hammerhead ribozyme. A kinetic description of the cleavage of matched and mismatched targets will be developed with the goal of predicting both the rate and specificity of the reaction. A strategy is proposed to identify sites in large RNAs that are particularly susceptible to cleavage. The inhibition of hammerhead cleavage by antibiotics will be explored. Finally, recently developed methods for in vitro evolution will be used to select for improved versions of the hammerhead and for new ribozymes that cleave pre-defined RNA targets.

该项目的目的是开发使用RNA酶作为试剂 对于现场特定的乳沟（随之而来的失活）， 其他高分子量RNA在体外和体内。 最近的成功 使用锤头核酶灭活许多RNA的几个实验室 （包括艾滋病毒序列）在细菌，植物和动物细胞中强调 这种新兴技术的价值。 这个项目将重点放在 精炼和扩展裂解的生化描述 锤头核酶大小的RNA。 动力学描述 将开发匹配和错配目标的分裂 预测反应的速率和特异性的目的。 一个 提出了策略来确定大型RNA中的站点 特别容易裂解。 锤头的抑制作用 将探索抗生素的切割。 最后，最近开发了 体外进化的方法将用于选择改进 锤头的版本和裂解的新核酶预先定义 RNA靶标。