DESIGN OF OPTIMAL RIBOZYMES FOR CLEAVAGE OF HIV RNA

本项目的目标是开发RNA酶作为试剂的用途 用于HIV的位点特异性切割（随后的失活）， 其他高分子量RNA在体外和体内。 最近成功地在 几个实验室使用锤头状核酶来复制一些RNA， （包括艾滋病毒序列）在细菌，植物和动物细胞中的作用， 这一新兴技术的价值。 该项目将重点关注 细化和扩展的生物化学描述的分裂 小的和大的RNA的锤头状核酶。 动力学描述 将开发匹配和不匹配目标的切割 目标是预测反应的速率和特异性。 一 提出了一种策略，以确定大RNA中的位点， 特别易受裂解的影响。 锤头的抑制 将探索通过抗生素的裂解。 最后，最近开发的 体外进化的方法将用于选择改进的 锤头的新版本和切割预定义的新核酶 RNA靶标。

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.