RNA MEDIATED GENETIC INTERFERENCE IN C ELEGANS

基因表达的靶向抑制是基础研究的重要工具。 医学研究在许多领域，并有许多潜在的临床 应用.然而，需要改进的工具来探测函数 克隆基因，特别是在考虑到全面的基因组 目前正在进行的许多生物体的测序项目。现时的建议 旨在理解和改进一种称为“RNAi”的工具， RNA介导的遗传干扰。RNA干扰是在 尝试使用“反义”方法来干扰基因 表达于C.优雅这些研究得出了令人惊讶的结论 “正义”或“反义”RNA链的制备可以诱导 有效的和特定的遗传干扰后显微注射到这个 有机体这项提案中描述的发现表明， 生物体的反应可能是这种遗传干扰的基础。值得注意的是， 不仅在整个组织中观察到干扰效应， 注射动物，但由100%的注射动物的遗传 后代在后代中，遗传干扰可以传递给 在精子或卵母细胞中作为显性基因外因子。微量注射 RNA（而不是DNA）可以诱导干扰。令人惊讶的是，双链 RNA在诱导干扰方面比任何一种单一的RNA都要有效得多。 搁浅。该提案概述了一种系统的方法，将剖析 RNAi的分子和遗传机制。目标将包括： 与干扰相关的属性。方法的改进。 并对C. elegans基因的项目介导遗传 干扰这些研究的见解可能直接导致 相关RNA干扰技术的发展 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.