Toll Signaling Pathway in Zebrafish Knockdowns

描述（由申请人提供）：本提案针对 Toll信号转导介导的天然免疫应答研究 通路通过检查Toll通路在响应于 传染病，并通过确定的方式，这方面的先天 免疫力可以调节适应性免疫力， 延长免疫反应为了检验每种蛋白质的作用 在Toll通路中，一种新的反义技术， 寡核苷酸（吗啉代寡核苷酸）阻断mRNA转录物的翻译， 将被用来生产斑马鱼击倒。吗啉代敲除 易于设计并且仅需要5'端未翻译的序列信息 非翻译区（UTR），翻译起始位点上游30 - 40个碱基 （8月）。这项新技术非常灵活， 下一代击倒鱼。此外，吗啉代翻译块 允许快速和容易地应用两种或三种寡核苷酸的组合， 产生击倒的斑马鱼。 该补助金的具体目标是： 1.为了产生Toll信号通路的吗啉代斑马鱼敲低， 2.用病毒和细菌性斑马鱼病原体来挑战击倒 通过检测斑马鱼Toll信号通路， 在保护性免疫对病原体的作用，我们将获得更完整的 了解先天免疫对免疫系统发育的影响 系统以及这条途径如何有助于形成一个 适应性免疫反应

DESCRIPTION (Provided by the Applicant): This proposal is directed toward the study of the innate immune response through the Toll signal transduction pathway. By examining the way in which Toll pathway functions in response to infectious diseases and by determining ways in which this facet of innate immunity may regulate adaptive immunity, it may be possible to augment, extend, and prolong the immune response. In order to examine the role of each protein in the Toll pathway, a new antisense technology, in which an alternative type of oligonucleotide (morpholino oligos) blocks translation of mRNA transcripts, will be used to produce zebrafish knock-downs. The morpholino knock-downs are easy to design and only require sequence information in the 5' untranslated region (UTR), thirty to forty bases upstream from the translational start site (AUG). This new technology is extremely flexible and will afford the rapid generation of knock-down fish. In addition, the morpholino translation block allows for quick and easy application of combinations of two or three oligos to generate knock-down zebrafish. The SPECIFIC AIMS for this grant are: 1. To generate morpholino zebrafish knock-downs of the Toll signaling pathway 2. To challenge the knock-downs with viral and bacterial zebrafish pathogens By examining the Toll signaling pathway in the zebrafish and determining its role in protective immunity against pathogens, we will gain a more complete understanding of the impact of innate immunity on the development of the immune system and how this pathway has helped to give rise to the formation of an adaptive immune response.