Development of a TILLING Resource for the Xenopus Research Community

为非洲爪蟾研究界开发 TILLING 资源

基本信息

批准号：
7945090
负责人：
Robert M Grainger
金额：
$ 25.19万
依托单位：
UNIVERSITY OF VIRGINIA
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-08-20 至 2012-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7945090
关键词：
Animal Housing Animal Model Animals Back Biological Biological Models Collaborations Communities Complement DNA DNA Sequence DNA Sequence Analysis Development Developmental Process Disease Dose Ethylnitrosourea Evolution Eye Development Funding Future Genes Genome Genomics Goals Grant Homeobox Genes Human Induced Mutation Informatics Label Laboratories Lesion Marines Methods Molecular Biology Mutagenesis Mutagens Mutate Mutation Phase Phenotype Population Preparation Protocols documentation Rana Regulator Genes Research Research Personnel Research Training Resources Sampling Scheme Sequence Analysis Site Spermatogonia Technology Wood material Work Xenopus animal resource base comparative ethylurea eye formation gene function high reward high risk insight interest male mutant next generation novel null mutation offspring public health relevance research study response sound sperm cell tool xenopus genome

项目摘要

DESCRIPTION (provided by applicant): During the past several years several model organism groups have used sequencing methods to screen through large populations of mutagenized animals (commonly referred to as TILLING) to identify mutations in known genes of high interest. We propose here to initiate development of a TILLING resource for the Xenopus research community at the Marine Biological Laboratory (MBL) at Woods Hole, a site chosen for two key reasons: 1) the impending construction of a national Xenopus Resource Center (XRC) there (for housing animals lines and advanced research training); and 2) the presence of an advanced sequencing facility associated with the highly regarded Josephine Bay Paul Center for Comparative Molecular Biology and Evolution at the MBL. The Grainger lab has already been involved in preparing animal samples for a small scale TILLING project at the Sanger Centre in the UK, where, using conventional sequencing, a very approximate hit rate for ENU mutagenesis has been determined. Recently the first null mutation has been identified in this screen, in the Rax gene (a key regulatory gene involved in eye formation), illustrating that the basic strategy is effective. In this proposal we have three goals. First we will use next generation sequencing to ascertain a far more accurate dose- response curve for ENU mutagenesis. Second, we will collect several thousand animals to provide material for a TILLING analysis. Third, we will refine next generation approaches for identifying mutations by TILLING, with the aim of identifying null mutations in several key regulatory genes. In addition to mutant animals generated in the Grainger lab, the Khokha and Conlon labs will also contribute samples to this effort. This project will provide the basis for development of a larger, long term TILLING project that will become part of the XRC, providing a resource for generating mutant lines for investigators nationwide. PUBLIC HEALTH RELEVANCE: Project Narrative The goal of this project is to identify mutations in genes important for the early development of the frog Xenopus. The genes under study, when mutated in humans, often result in genetically based diseases, and study of these mutations in the frog will provide new insights about these diseases because of the unique tools available to researchers for clarifying gene function in these animals.

描述（由申请人提供）：在过去的几年中，几个模型有机体群体已经使用了测序方法来筛选大量诱变动物种群（通常称为耕种），以鉴定已知的高兴趣基因中的突变。我们在这里建议在伍兹霍尔（Woods Hole）的海洋生物实验室（MBL）的Xenopus研究社区开发耕作资源，该地点是出于两个关键原因：1）即将在那里建造国家Xenopus资源中心（XRC）（用于住房动物线和高级研究培训）； 2）与高度认可的约瑟芬湾保罗比较分子生物学和MBL相关的高级测序设施的存在。 Grainger实验室已经参与了在英国Sanger中心为小规模耕作项目准备动物样品，在那里，使用常规测序，已经确定了ENU诱变的近似命中率。最近，在RAX基因（与眼睛形成有关的关键调节基因）中已经确定了第一个无效突变，这说明了基本策略是有效的。在此提案中，我们有三个目标。首先，我们将使用下一代测序来确定ENU诱变的更准确的剂量响应曲线。其次，我们将收集数千只动物，以提供耕种分析的材料。第三，我们将完善下一代方法来通过耕种来识别突变，目的是识别几个关键调节基因中的无效突变。除了在Grainger实验室中产生的突变动物外，Khokha和Conlon Labs还将为这项工作贡献样本。该项目将为开发更大的长期耕作项目的发展提供基础，该项目将成为XRC的一部分，为全国调查人员生成突变线的资源提供了资源。公共卫生相关性：项目叙述该项目的目的是确定基因中对青蛙爪蟾早期发展重要的基因突变。研究的基因在人类中发生突变时通常会导致基于遗传的疾病，并且对青蛙中的这些突变的研究将提供有关这些疾病的新见解，因为研究人员可以使用独特的工具来阐明这些动物中的基因功能。