RNA MEDIATED GENETIC INTERFERENCE IN C ELEGANS

靶向抑制基因表达是基础研究的必要工具 医学研究涉及多个领域，具有许多临床潜力 申请。然而，需要改进的工具来探测这些功能 克隆的基因，特别是在综合基因组的情况下 许多生物体的测序项目目前正在进行中。目前的建议 旨在理解和改进一种称为“RNAi”的工具 用于RNA介导的遗传干扰。RNAi是在这个过程中发现的 企图使用“反义”方法干扰基因 在线虫中的表达。这些研究导致了令人惊讶的观察结果 无论是“正义”还是“反义”的RNA链的制备都可以诱导 对此进行显微注射时的有效和特异的遗传干扰 有机体。这项提案中描述的发现表明，细胞和 生物体的反应可能是这种基因干扰的基础。值得注意的是， 干扰效应不仅在整个组织中观察到，而且 被注射的动物，但100%被注射的动物的 后代。在以后的世代中，遗传干扰可以传播 在精子或卵母细胞中作为主要的基因外因素。微量注射 RNA(但不是DNA)可以引起干扰。令人惊讶的是，双重搁浅 RNA在诱导干扰方面比单独的任何一个都要有效得多 斯特兰德。这份提案概述了一种系统的方法，它将剖析 RNAi背后的分子和遗传机制。AIMS将包括： 与碰撞关联的特性。方法上的改进。 并鉴定其投影介导遗传的线虫基因 干扰。来自这些研究的见解可能会直接导致 基因探测相关RNA干扰技术的研究进展 在包括人类在内的其他生物体中发挥作用。

The targeted inhibition of gene expression is an essential tool for basic medical research in numerous fields and has many potential clinical applications. However, improved tools are needed for probing the functions of cloned genes, especially in light of the comprehensive genome sequencing projects now underway for many organisms. The present proposal is directed at understanding and improving one such tool known as "RNAi" for RNA mediated genetic interference. RNAI was discovered in the course of attempts to use "antisense" methodology to interfere with gene expression in C. elegans. These studies led to the surprising observation that preparations of either "sense" or "antisense" RNA strands can induce potent and specific genetic interference upon microinjection into this organism. Findings described in this proposal suggest that a cellular and organismal response may underlie this genetic interference. Remarkably, interfering effects are observed not only throughout the tissues of the injected animal but are inherited by 100% of the injected animal's progeny. In subsequent generations genetic interference can be transmitted in the sperm or oocytes as a dominant extragenic factor. Microinjection of RNA (but not DNA) can induce interference. Surprisingly, double stranded RNA is much more effective at inducing interference than is either single strand. This proposal outlines a systematic approach that will dissect the molecular and genetic mechanisms that underlie RNAi. Aims will include: properties associated with interference. Improvements in the methodology. And identification of the C. elegans genes whose projects mediate genetic interference. Insights from these studies may lead directly to the development of related RNA interference technologies for probing gene function in other organisms including humans.