Functional Genomics in a Simple Model Metazoan

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，并研究重要的新后生动物模型系统Nematostella vectensis胚胎中的基因功能。 新的研究结果来自于N. vectensis表明脊椎动物与刺胞动物如刺胞动物在基因组内容上有更大的相似性。vectens比任何一个苍蝇或线虫。 这一发现使人们对衍生的遗传模型系统可能对人类生物学的意义产生怀疑，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。 N. vectensis将允许理解在脊椎动物中看到的发育模式的分子控制的差异，但在衍生的蜕皮动物遗传模型系统中没有，提供了对脊椎动物谱系辐射之前部署的古老分子模式机制的重要见解。 能够在这些动物中实验性地剖析分子通路，对于理解正常人类健康和疾病所涉及的遗传调控网络的变异起源至关重要。 这项工作利用了N. vectensis由联合基因组研究所（能源部），并将提供额外的技术和方法背景，为今后的实验领域。 这一申请将大大提高我们的能力，启动一个长期的研究计划，基因发现和功能调查的遗传调控途径共同所有活着的动物。 本申请的目的是：具体目的1：优化转基因应用的电穿孔递送技术。 具体目标2：利用转座子介导的插入技术创建稳定的转基因动物。 具体目标3：产生表达通过BAC克隆的同源重组标记的基因座的转基因动物。 表达构建体已经产生，并且坚实的初步数据沿着具有证明成就历史的人员，确保将取得快速进展。 这些数据将提供重要的发育事件，如原肠胚形成过程中的胚层形成，双边对称性的起源，神经发育/组织，和体细胞/生殖干细胞生物学的增强子陷阱技术和插入突变的基因网络进化的功能分析的基因发现的技术基础。

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.