Zebrafish Mutant Mapping Facility

斑马鱼突变体绘图设施

基本信息

批准号：
7267020
负责人：
RONALD G GREGG
金额：
$ 20.91万
依托单位：
UNIVERSITY OF LOUISVILLE
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-08-01 至 2009-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7267020
关键词：
Animal Model Biochemical Pathway Candidate Disease Gene Chemicals Chromosome Mapping Chromosomes Commit Computer Retrieval of Information on Scientific Projects Database Core Facility DNA Data Databases Disease Dyes Embryo Event Fees Fluorescence Funding Genes Genetic Genetic Polymorphism Genetic Recombination Genome Genotype Goals Grant Individual Inheritance Patterns Internet Label Laboratories Length Link Location Maps Modeling Molecular Mutagenesis Mutation Nature Numbers Parents Phase Phenotype Polymerase Chain Reaction Publications Publishing Range Rate Reagent Recombinants Research Personnel Resolution Resources Rights Screening procedure Services Site Staging Standards of Weights and Measures Technical Expertise Time Tissues Transfer Agreement United States National Institutes of Health Zebrafish base cost day gene discovery genome sequencing human disease insight interest male mutant novel positional cloning programs response size tool

项目摘要

DESCRIPTION (provided by applicant): The objective of this proposal is to accelerate the identification of zebrafish genes discovered during mutagenesis screens. This proposal is in response to PAR-02-142 to develop "Tools for Genetic Studies in Zebrafish." A major focus of this PA is to use novel screens to identify mutants of interest. However, an essential part of this effort is to identify the genes responsible for the novel phenotypes generated. While thousands of mutants already have been described, only 167 genes are listed in the zebrafish database (ZFIN) for which mutations are known (although there are probably some not listed). The major reason for this is the time consuming nature of the positional cloning strategy that must be used. The major bottleneck in gene discovery by positional cloning is the need to do whole genome linkage mapping. This takes considerable resources and time, as well as technical expertise. This goal of this proposal is to eliminate this bottleneck by setting up a high throughput mapping facility. The usual laborious task of screening sometimes hundreds of markers to find one that is linked to the mutation of interest will be accomplished in 2 days. This will be done using fluorescence sequencers to automate genotyping of SSLPs (short sequence length polymorphisms) that span the genome. The mapping will be done in two main stages. First, an approximate location will be determined using bulk segregant analyses. Second, once closely flanking markers are identified 500-1000 individual mutant embryos will be genotyped to identify the region containing the mutant gene. Third, for a small number of mutants the genes will be identified. The core will map a minimum of 100 mutants per year and identify the gene in 5-10, although both these numbers are likely to increase dramatically once the genome sequence is completed. The service will be modular so that investigators can have the initial mapping phases done by the facility, and then complete the mutant gene isolation in there own laboratories. This project will have considerable impact because it will greatly accelerate the discovery of genes that already have been shown to be models of human diseases. The facility is committed to full and open access and all data pertaining to mapping will be made available at publication. Given the importance of zebrafish as a model organism this facility will have an immediate impact on the rate at which mutant genes are discovered, which will provide new insights into the genetic underpinnings of important biochemical pathways involved in disease.

描述（由申请人提供）：该提案的目的是加速对诱变筛选过程中发现的斑马鱼基因的鉴定。该提案是为了响应 PAR-02-142，开发“斑马鱼遗传研究工具”。该 PA 的一个主要重点是使用新颖的屏幕来识别感兴趣的突变体。然而，这项工作的一个重要部分是确定负责产生新表型的基因。虽然已经描述了数千个突变体，但斑马鱼数据库 (ZFIN) 中仅列出了已知突变的 167 个基因（尽管可能有一些未列出）。其主要原因是必须使用的定位克隆策略非常耗时。通过定位克隆发现基因的主要瓶颈是需要进行全基因组连锁图谱。这需要大量的资源和时间以及技术专业知识。该提案的目标是通过建立高吞吐量测绘设施来消除这一瓶颈。通常需要筛选数百个标记以找到与目标突变相关的标记，这一艰巨的任务将在 2 天内完成。这将使用荧光测序仪对跨越基因组的 SSLP（短序列长度多态性）进行自动化基因分型来完成。映射将分两个主要阶段完成。首先，将使用批量分离分析来确定大致位置。其次，一旦鉴定出紧邻的标记，将对 500-1000 个单独的突变胚胎进行基因分型，以鉴定含有突变基因的区域。第三，对于少数突变体，将鉴定基因。核心每年将绘制至少 100 个突变体图谱，并在 5-10 个突变体中识别出基因，尽管一旦基因组序列完成，这两个数字可能会急剧增加。该服务将是模块化的，以便研究人员可以由该设施完成初始绘图阶段，然后在自己的实验室中完成突变基因分离。该项目将产生相当大的影响，因为它将大大加速已被证明是人类疾病模型的基因的发现。该设施致力于完全开放的访问，所有与测绘相关的数据将在发布时提供。鉴于斑马鱼作为模式生物的重要性，该设施将对突变基因的发现速度产生直接影响，这将为疾病相关重要生化途径的遗传基础提供新的见解。

项目成果

期刊论文数量（7）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Mechanisms underlying metabolic and neural defects in zebrafish and human multiple acyl-CoA dehydrogenase deficiency (MADD).

DOI：
10.1371/journal.pone.0008329
发表时间：
2009-12-17
期刊：
PloS one
影响因子：
3.7
作者：
Song Y;Selak MA;Watson CT;Coutts C;Scherer PC;Panzer JA;Gibbs S;Scott MO;Willer G;Gregg RG;Ali DW;Bennett MJ;Balice-Gordon RJ
通讯作者：
Balice-Gordon RJ

Loss of Lrp2 in zebrafish disrupts pronephric tubular clearance but not forebrain development.

DOI：
10.1002/dvdy.22624
发表时间：
2011-06
期刊：
DEVELOPMENTAL DYNAMICS
影响因子：
2.5
作者：
Kur, Esther;Christa, Anna;Veth, Kerry N.;Gajera, Chandresh R.;Andrade-Navarro, Miguel A.;Zhang, Jingjing;Willer, Jason R.;Gregg, Ronald G.;Abdelilah-Seyfried, Salim;Bachmann, Sebastian;Link, Brian A.;Hammes, Annette;Willnow, Thomas E.
通讯作者：
Willnow, Thomas E.

Genetic Dissection of Dual Roles for the Transcription Factor six7 in Photoreceptor Development and Patterning in Zebrafish.