ZEBRAFISH GENES INVOLVED IN EARLY EMBRYONIC DEVELOPMENT

参与早期胚胎发育的斑马鱼基因

• 批准号: 3329261
• 负责人: GARY W STUART
• 金额: $ 8.55万
• 依托单位: INDIANA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-03-01 至 1996-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3329261
• 关键词: complementary DNA; cytogenetics; developmental genetics; early embryonic stage; embryogenesis; enzyme mechanism; gene expression; gene mutation; genetic enhancer element; genetic markers; genetic models; genetic promoter element; genetically modified animals; germ cells; haploidy; in situ hybridization; laboratory mouse; microinjections; molecular cloning; northern blottings; nucleic acid probes; nucleic acid sequence; tissue /cell culture; transfection; vertebrate embryology; zebrafish

In order to understand the basis of human genetic disease and to begin to develop procedures by which human developmental anomalies can be treated, it is necessary to develop appropriate animal models and to perform basic research designed to uncover the molecular mechanisms of normal and abnormal development. Progress toward a molecular-genetic description of the embryonic development of invertebrate organisms such as D. melanogaster and C. elegans has been aided by their short generation times. To elucidate the molecular mechanisms by which vertebrate embryos develop, a similar analysis is required. However, inquiries into the molecular-genetic basis of vertebrate development have been hindered by long generation times, and in the case of mammals, inaccessible embryos (i.e., in utero development). In the investigations proposed here, a molecular-genetic analysis of early vertebrate development will be initiated using the zebrafish as a model system. This organism has many experimental advantages relative to mammalian systems, including small size, fecundity, and the natural in vitro development of optically transparent embryos. In addition, it is possible to produce and analyze gynogenetic and haploid progeny, and to produce stable germ-line transformants. Genetic loci likely to be involved in early embryonic development will be identified with the aid of a transgene marker designed to respond to endogenous enhancer-like controlling elements located near the site of integration. The unique expression patterns exhibited by these integrated "enhancer detectors" will provide important clues as to the function of the marked genes. The potential of these transgenes to result in insertion mutations with interesting phenotypes will also be evaluated. The transgenes themselves provide hybridization and/or genetic markers allowing the rapid recovery of flanking sequences at the identified locus. Marked loci will be further characterized by Southern and Northern analysis, allowing the identification of individual genes likely to play crucial roles in development.

为了了解人类遗传疾病的基础并开始 制定治疗人类发育异常的程序， 有必要开发适当的动物模型并进行基础研究 研究旨在揭示正常和 发育异常。分子遗传学描述的进展 无脊椎动物（如黑腹果蝇）的胚胎发育 秀丽隐杆线虫的世代时间较短，这也为线虫提供了帮助。阐明 脊椎动物胚胎发育的分子机制，类似 需要进行分析。然而，对分子遗传学基础的探究 脊椎动物的发育受到长世代时间的阻碍，并且 对于哺乳动物来说，无法获得胚胎（即在子宫内发育）。 在这里提出的研究中，对早期的分子遗传学分析 脊椎动物的发育将以斑马鱼为模型开始 系统。相对于该生物体，该生物体具有许多实验优势 哺乳动物系统，包括体型小、繁殖力强和自然环境 光学透明胚胎的体外发育。此外，它是 可以产生和分析雌核发育和单倍体后代，并且 产生稳定的种系转化体。可能涉及的遗传位点 在早期胚胎发育中，将借助 旨在响应内源增强子样的转基因标记 控制元件位于集成位置附近。独特的 这些集成的“增强子检测器”表现出的表达模式将 为标记基因的功能提供重要线索。这 这些转基因可能导致插入突变 还将评估有趣的表型。转基因本身 提供杂交和/或遗传标记，允许快速恢复 已识别基因座的侧翼序列。标记的位点将进一步 以南方和北方分析为特征，允许 鉴定可能发挥关键作用的个体基因 发展。