Discovery Craniofacial and Retinal Genes

描述（由申请人提供）：为了响应RFA DE-03-001“颅面疾病的基因发现”，本申请是为了在斑马鱼中进行有效基因发现的独特计划，以发现人类疾病基因的候选者：我们的具体目标是发现斑马鱼基因，当突变时，这些基因会导致（1）颅面骨发育不良和（2）视网膜变性。对于这两个，一个新的强大的基因发现方法的组合将被利用。基因中的损伤将通过病毒介导的插入诱变引入。由于病毒插入物可以标记突变的基因，因此可以非常迅速地识别和克隆基因，通常在几天内。突变体将在高效、定向、第一代（G1）筛选中鉴定，使用通过孤雌生殖获得的胚胎和幼虫，并用荧光染料进行活体染色以揭示表型。孤雌生殖和G1筛选消除了几个月的时间，并节省了饲养数十万条鱼所需的更常见的前瞻性遗传策略，提高家庭的突变体和筛选F3后代。新的菌株将迅速提供给其他研究人员使用社区资源，斑马鱼信息网络宣传突变体和斑马鱼国际资源中心分发它们。 斑马鱼胚胎和幼体的各种有利属性将允许详细的表型研究和快速表征基因在面骨和眼睛发育中的功能。

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.