Discovery Craniofacial and Retinal Genes

发现颅面和视网膜基因

• 批准号: 6752818
• 负责人: CHARLES B KIMMEL
• 金额: $ 32.7万
• 依托单位: UNIVERSITY OF OREGON
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-06-01 至 2006-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6752818
• 关键词: animal genetic material tag; biotechnology; craniofacial; developmental genetics; fluorescent dye /probe; gene expression; gene mutation; genetic screening; in situ hybridization; molecular cloning; parthenogenesis; phenotype; polymerase chain reaction; retina degeneration; skeletal disorder; zebrafish

DESCRIPTION (provided by applicant): In response to RFA DE-03-001 "Gene Discovery for Craniofacial Disorders" this application is for a unique program of efficient gene discovery in zebrafish to find candidates for human disease genes: Our specific aims are to discover zebrafish genes that when mutated result in (1) craniofacial bone dysgenesis, and (2) retinal degeneration. For both, a novel combination of powerful approaches toward gene discovery will be utilized. Lesions in genes will be introduced by viral-mediated insertional mutagenesis. Because the viral insertions serve to tag the mutated genes, the genes can be identified and cloned extremely rapidly, often within a few days. The mutants will be identified in highly efficient, directed, first generation (G1) screens, using embryos and larvae obtained by parthenogenesis and vitally stained with fluorescent dyes to reveal the phenotypes. Parthenogenesis and G1 screening eliminates months of time and saves rearing hundreds of thousands of fish required in the more usual forward-genetic strategies of raising families of mutants and screening F3 progeny. The new strains will be made rapidly available to other researchers using community resources, the Zebrafish Information Network to advertise the mutants and the Zebrafish International Resource Center to distribute them. Various favorable attributes of the zebrafish embryo and larva will permit detailed phenotypic studies and rapid characterization of the functions of the genes in facial bone and eye development.

描述（由申请人提供）：响应RFA DE-03-001“颅面疾病的基因发现基因发现”该应用是针对斑马鱼中有效基因发现的独特程序，以找到人类疾病基因的候选者：我们的特定目的是发现发现斑马鱼基因在突变后会导致（1）颅骨失调，以及（2）视网膜变性。对于这两者来说，将利用强大的基因发现方法的新型组合。基因的病变将由病毒介导的插入诱变引入。由于病毒插入可用于标记突变基因，因此通常在几天之内可以非常快速地识别和克隆基因。该突变体将在高效的，有向的，第一代（G1）筛选中鉴定，使用单骨生成获得，并用荧光染料进行了性染色以揭示表型。 par骨生成和G1筛查消除了数月的时间，并节省了饲养数十万条鱼类在养育突变体家族和筛选F3后代的更常见的前向遗传策略中所需的数十万条鱼。新的菌株将使用社区资源，斑马鱼信息网络迅速向其他研究人员提供，以宣传突变体和斑马鱼国际资源中心，以分发它们。 斑马鱼胚胎和幼虫的各种有利属性将允许详细的表型研究以及对面部骨骼和眼睛发育中基因功能的快速表征。