Development of in Vitro Genetic System to Study RNA Editing

研究RNA编辑的体外遗传系统的开发

• 批准号: 9520253
• 负责人: Laura Landweber
• 金额: $ 5.3万
• 依托单位: Princeton University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-04-01 至 1997-03-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9520253&HistoricalAwards=false
• 关键词: Development; Vitro; Genetic; System; Study

9520253 Landweber The objective of this research is to develop the use of in vitro selection as a model system to explore he possible link between RNA catalysis and kinetoplastid RNA editing. RNA editing is the bizarre and complex addition or deletion of uridine residues which occurs in the mitochondria of some of the earliest eukaryotes, the kinetoplastid protozoa. The approach will be to develop a successful in vitro selection system to analyze the mechanism and evolution of RNA editing. These experiments will address the antiquity of RNA editing and the role of RNA catalysts in evolution. One hypothesis is that RNA editing is an ancient process, and the interaction of several short loci to produce one final transcript may reflect some of the steps involved in the first assembly of genes. Specifically, these experiments will test the hypothesis that RNA editing may have been one of the key players in the RNA world by analyzing whether each of its steps can be catalyzed by RNA, a characteristic of some of the earliest hypothesized biological mechanisms. This research will determine whether the first step of RNA editing, a transesterification similar to other RNA catalyzed ligation reactions, can be catalyzed by an RNA molecule. Through a series of in vitro selection experiments, which simulate evolution in a test tube, the experiments will explore the world of possible primitive RNA editing enzymes, by selecting the desired catalytic molecules from a pool of random sequences. Armed with these preliminary experiments, we will then be prepared to analyze the later steps of RNA editing, with the final goal of evolving a complete RNA catalyzed RNA editing system in vitro. These experiments will test whether a molecule capable of editing RNA could have arisen under fairly simple conditions: a model for the evolution of complex molecular processes. Ultimately, this will allow one to design new RNA enzymes that can edit RNA of any specific sequence. %%% The objective od this research is to deve lop the use of in vitro selection to explore the possible link between RNA catalysis and kinetoplastid RNA editing. RNA editing is the bizarre and complex addition or deletion of uridine residues which occurs in the mitochondria of some of the earliest eukaryotes, the kinetoplastid protozoa. The approach will be to develop a successful in vitro selection system to analyze the mechanism and evolution of RNA editing. These experiments will test whether a molecule capable of editing RNA could have arisen under fairly simple conditions: a model for the evolution of complex molecular processes. Through a series of in vitro selection experiments, which simulate evolution in a test tube, the experiments will explore the world of possible primitive RNA editing enzymes, by seleating new catalytic molecules from a pool of random sequences. Ultimately, this will allow one to design new RNA enzymes that can edit RNA of any specific sequence. ***

9520253 Landweber本研究的目的是开发体外选择作为模型系统的用途，以探索RNA催化和动质体RNA编辑之间可能的联系。RNA编辑是尿苷残基的奇异而复杂的添加或缺失，其发生在一些最早的真核生物（动质体原生动物）的线粒体中。该方法将开发一个成功的体外选择系统来分析RNA编辑的机制和进化。 这些实验将解决RNA编辑的古老性和RNA催化剂在进化中的作用。一种假设是RNA编辑是一个古老的过程，几个短基因座的相互作用产生一个最终的转录本可能反映了基因首次组装的一些步骤。具体来说，这些实验将通过分析RNA编辑的每个步骤是否可以被RNA催化来验证RNA编辑可能是RNA世界中的关键角色之一的假设，这是一些最早假设的生物机制的特征。这项研究将确定RNA编辑的第一步，类似于其他RNA催化的连接反应的酯交换，是否可以由RNA分子催化。 通过一系列模拟试管进化的体外选择实验，这些实验将通过从随机序列库中选择所需的催化分子来探索可能的原始RNA编辑酶的世界。 有了这些初步的实验，我们将准备分析RNA编辑的后续步骤，最终目标是在体外开发一个完整的RNA催化RNA编辑系统。这些实验将测试一种能够编辑RNA的分子是否可以在相当简单的条件下出现：复杂分子过程进化的模型。最终，这将允许设计新的RNA酶，可以编辑任何特定序列的RNA。 本研究的目的是开发体外选择的使用，以探索RNA催化和动质体RNA编辑之间的可能联系。RNA编辑是尿苷残基的奇异而复杂的添加或缺失，其发生在一些最早的真核生物（动质体原生动物）的线粒体中。该方法将开发一个成功的体外选择系统来分析RNA编辑的机制和进化。 这些实验将测试一种能够编辑RNA的分子是否可以在相当简单的条件下出现：复杂分子过程进化的模型。通过一系列模拟试管进化的体外选择实验，这些实验将通过从随机序列库中选择新的催化分子来探索可能的原始RNA编辑酶的世界。最终，这将允许设计新的RNA酶，可以编辑任何特定序列的RNA。 ***