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: 3145295
• 负责人: OLKE C UHLENBECK
• 金额: $ 10.95万
• 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 1993-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3145295
• 关键词: AIDS; AIDS therapy; HIV infections; antisense nucleic acid; chemical binding; conformation; drug delivery systems; endonuclease; enzyme structure; gel electrophoresis; human immunodeficiency virus; immunotherapy; molecular genetics; molecular site; nucleic acid denaturation; nucleic acid hybridization; nucleic acid sequence; nucleic acid structure; polymerase chain reaction; protein engineering; radionuclides; reagent /indicator; ribozymes; virus RNA

The goal of this project is to understand the biochemical properties of the "hammerhead" and "hairpin" RNA enzymes (ribozymes). These two small RNA catalysts have the potential to cause site-specific cleavage (and consequent inactivation) of HIV and other high molecular weight RNAs in vitro and in vivo. The major focus will be on the design of optimally active cleavage reagents in vitro. Important variables include the possibility of interfering structure in the target RNA, the conformational homogeneity of the ribozyme and target RNAs, the length and sequence of the RNA helices which join the RNA enzyme to the target and the possibility of certain RNA structures which stimulate hybridization rates. The approach will involve the use of pre-steady state and steady state kinetics to analyze the cleavage reaction. The rate limiting step of each reaction will be determined since it is anticipated that it may change depending on reaction conditions. If possible,, the Kappa/m will be related to the known free energy parameters of helix formation, making it possible to predict the Kappa/m for any ribozyme-substrate pair.

该项目的目的是了解 “ Hammerhead”和“发夹” RNA酶（核酶）。 这两个小RNA 催化剂有可能引起特定地点的切割（和 随之而来的艾滋病毒和其他高分子量RNA的失活） 体内和体内。 主要重点将放在最佳设计上 活性切割试剂体外。 重要变量包括 在靶RNA中干扰结构的可能性，构象 核酶和靶RNA的均匀性，该长度和序列 将RNA酶连接到靶的RNA螺旋以及可能性 刺激杂交速率的某些RNA结构。 该方法将涉及使用前态状态和稳定状态 动力学分析裂解反应。 每个的速率限制步骤 将确定反应，因为预计可能会改变 取决于反应条件。 如果可能的话，kappa/m将是 与螺旋形成的已知自由能参数有关，使其 可以预测任何核酶 - 基底对的kappa/m。