Development of RNA interference in zebrafish

描述(申请人提供)：功能基因组学需要可靠的方法来研究基因功能中断的后果。自从在线虫中发现以来，RNA干扰(RNAi)已经成为在细胞培养、哺乳动物和无脊椎动物模型系统中处理基因功能的宝贵资源。然而，事实证明，将这项技术应用于斑马鱼和非洲爪哇是一个重大挑战。在这项提案中，我们将应用我们在microRNA(MiRNA)领域的现有知识来开发几种改进RNAi的策略。这项提议将向科学界提供具体规则，以改进斑马鱼的RNAi，以便能够对斑马鱼基因的功能进行表征。在未来，该项目的结果将建立一个框架，以了解脊椎动物基因的结构和功能。 公共卫生相关性：了解基因的功能需要可靠的方法来阻断基因表达。在过去的十年里，人们发现双链RNA可以用来关闭基因的表达(RNA干扰)，但这种技术在斑马鱼中是不可用的。这项建议旨在了解将这项强大的技术应用于斑马鱼所需的步骤，这将有助于我们了解脊椎动物基因在发育和疾病中的功能。

DESCRIPTION (provided by applicant): Functional genomics requires reliable methods to investigate the consequences of disrupting the gene function. Since its discovery in C .elegans, RNA interference (RNAi) has become an invaluable resource to tackle gene function in cell culture, mammals and invertebrate model systems. However, but it has proven a major challenge to apply this technique in zebrafish and Xenopus. In this proposal we will apply our current knowledge in the microRNA (miRNA) field to develop several strategies that improve RNAi. This proposal will provide specific rules to the scientific community to improve RNAi in zebrafish to allow the functional characterization of the zebrafish genes. In the future, the resuls derived from this project will establish a frame work to understand the structure and the function of vertebrate genes. PUBLIC HEALTH RELEVANCE: Understanding the function of genes requires reliable methods to block gene expression. Over the last decade, it has been discovered that double strand RNA can be used to shut down expression of genes (RNA interference) however this technique is not available in zebrafish. This proposal aims to understand the steps required to apply this powerful technique to zebrafish, what will help us understand the function of vertebrate genes in development and disease.