Functional Genomics in a Simple Model Metazoan

简单模型后生动物的功能基因组学

• 批准号: 7309580
• 负责人: MARK q MARTINDALE
• 金额: $ 6.92万
• 依托单位: UNIVERSITY OF HAWAII AT MANOA
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-20 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7309580
• 关键词: Animals; Bilateral; Biological Models; Cellular biology; Cnidaria; Data; Department of Energy; Development; Disease; Electroporation; Embryo; Enhancers; Event; Evolution; Expressed Sequence Tags; Future; Gene Expression; Genes; Genetic; Genetic Variation; Genome; Germ; Germ Layers; Goals; Health; Human; Human Biology; Human Resources; Insertional Mutagenesis; Institutes; Investigation; Joints; Label; Life; Mediating; Microinjections; Modeling; Molecular; Nematoda; Nuclear; Nucleic Acids; Pathway interactions; Pattern; Phylogenetic Analysis; Positioning Attribute; Procedures; Radiation; Recording of previous events; Regulatory Pathway; Reporter; Research; Solid; Stem cells; Techniques; Technology; Testing; Transgenes; Transgenic Animals; Uncertainty; Variant; Vertebrates; Work; base; cell type; fly; functional genomics; gene discovery; gene function; genome sequencing; homologous recombination; insight; neurodevelopment; programs; research study; tool; vertebrate genome

DESCRIPTION (provided by applicant): This application will develop robust techniques to manipulate gene expression and investigate gene function in embryos of an important new metazoan model system, Nematostella vectensis. Surprising new results from total genome sequencing and EST analyses of N. vectensis indicate that vertebrates share greater similarities in genome content with cnidarians like N. vectensis than either do with flies or nematodes. This revelation casts doubt on the significance that derived genetic model systems may have on human biology and thus provides the opportunity to understand the ancestral relationships between genetic variation and morphological complexity. The crucial phylogenetic position of N. vectensis will allow an understanding of the differences in the molecular control of developmental patterning seen in vertebrates, but absent in derived ecdysozoan genetic models systems, providing important insight into the ancient molecular patterning mechanism deployed before the radiation of vertebrate lineages. Being able to experimentally dissect molecular pathways in these animals is crucial to understanding the origin of variation in genetic regulatory networks involved in normal human health and disease. This work takes advantage of the complete genome sequencing of N. vectensis by the Joint Genome Institute (Department of Energy) and will provide additional technical and methodological background for future experiments in the field. This application will substantially enhance our ability to launch a long term research program for gene discovery and functional investigations of genetic regulatory pathways common to all living animals. The aims of this application are: Specific Aim 1: to optimize electroporation delivery techniques for transgene applications. Specific Aim 2: to create stable transgenic animals using transposon mediated insertion technology. Specific Aim 3: generate transgenic animals expressing loci marked through homologous recombination of BAC clones. Expression constructs have already been generated and solid preliminary data along with personnel with proven history of accomplishment insure that rapid progress will be made. These data will provide the technical basis for gene discovery for important developmental events such as germ layer formation during gastrulation, origin of bilateral symmetry, neural development/organization, and somatic/germ stem cell biology by enhancer trap techniques and for the functional analysis of gene network evolution by insertional mutagenesis.

描述（由申请人提供）：本申请将开发强大的技术来操纵基因表达，并研究一个重要的新的后生动物模型系统-线虫病的胚胎中的基因功能。来自全基因组测序和EST分析的令人惊讶的新结果表明，脊椎动物与线虫等刺胞动物的基因组内容比与苍蝇或线虫的相似性更大。这一发现使人们对衍生遗传模型系统在人类生物学上的重要性产生了怀疑，从而为理解遗传变异和形态复杂性之间的祖先关系提供了机会。vectensis的关键系统发育位置将使我们能够理解在脊椎动物中看到的发育模式的分子控制差异，但在衍生的外生动物遗传模型系统中却没有，这为在脊椎动物谱系辐射之前部署的古老分子模式机制提供了重要的见解。能够通过实验解剖这些动物的分子通路对于理解涉及正常人类健康和疾病的遗传调控网络变异的起源至关重要。这项工作利用了联合基因组研究所（能源部）对贝氏螺旋体的全基因组测序，并将为该领域未来的实验提供额外的技术和方法背景。这个应用程序将大大提高我们启动一个长期研究项目的能力，用于基因发现和所有现存动物共同的基因调控途径的功能调查。本申请的目的是：具体目标1：优化转基因应用的电穿孔递送技术。具体目标2：利用转座子介导插入技术建立稳定的转基因动物。特异性目的3：通过同源重组BAC克隆产生表达基因座的转基因动物。表达结构已经产生，坚实的初步数据以及具有证明的成就历史的人员确保将取得快速进展。这些数据将为通过增强子陷阱技术发现重要发育事件的基因提供技术基础，如原肠胚形成过程中胚层的形成、双边对称的起源、神经发育/组织和体细胞/生殖干细胞生物学，以及通过插入诱变技术分析基因网络进化的功能。