Gene Manipulation of Cephalopod Development

头足类动物发育的基因操作

• 批准号: 7614267
• 负责人: Clifton W. Ragsdale
• 金额: $ 7.68万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-05-01 至 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7614267
• 关键词: Alkaline Phosphatase; Animal Model; Animals; Biological; Biological Models; Biology; Blastoderm; Brain; Chemicals; Cuttlefish; Data; Development; Drosophila melanogaster; Electroporation; Embryo; Embryology; Future; Gene Expression; Gene Transfer; Gene Transfer Techniques; Genes; Genetic; Germ Lines; Goals; Human; Human Development; In Situ Hybridization; Invertebrates; Investments; Mediating; Methods; Modeling; Molecular; Mus; Nervous system structure; Octopus; Organism; Organogenesis; Plasmids; Polymers; Preparation; Promoter Regions; Reagent; Research; Research Personnel; Resources; Squid; Staging; System; Testing; Tissues; Tomatoes; Ubiquitin C; Vertebral column; base; embryo tissue; expression vector; hatching; innovation; insight; marine organism; plasmid DNA; promoter; vector

DESCRIPTION (provided by applicant): Understanding normal human development depends on animal model systems for experimental insight into underlying biological mechanisms. The long-term goal of this project is to use cephalopod biology to understand human organogenesis, including brain development. Among animals, human beings are large organisms with disproportionately large brains. Cephalopods, which include squids, octopuses, and cuttlefish, are the largest non-mammalian marine organisms and have the largest brains among all invertebrates. To study cephalopod development at a mechanistic level requires the identification of a cephalopod with ready availability, easy husbandry, and an embryology permitting experimental manipulation. The investigators have identified such a preparation, the Californian mud-flat octopus, O. bimaculoides. To date, they have developed methods for ex ovo culturing and for whole mount in situ hybridization for embryonic stages from the blastoderm to hatching. The investigators have also isolated a large panel of cDNAs for key developmental control genes. To complete this model system, they need to develop reliable methods of gene manipulation in these cephalopod embryos. Specific Aim 1 is to generate cephalopod-specific expression vectors based on octopus EF1-a and ubiquitin C genes. Specific Aim 2 is to develop methods of plasmid delivery to octopus embryos, for which the investigators will test as methods electroporation and chemical transgenesis using cationic polymers. Development of gene manipulation resources and methods will permit hypothesis-driven research of the molecular and cellular mechanisms of cephalopod development.

描述（由申请人提供）：理解正常的人类发展取决于动物模型系统，以实验对潜在生物学机制的实验洞察力。该项目的长期目标是使用头足类生物学来了解人体器官发生，包括大脑发育。在动物中，人类是大脑不成比例的大生物。包括鱿鱼，章鱼和墨鱼在内的头足动物是最大的非哺乳动物海洋生物，并且在所有无脊椎动物中的大脑中最大。要在机械水平上研究头足动物的发展，需要鉴定具有可用性，易用的饲养和允许实验性操作的胚胎学的头足类动物。研究人员已经确定了这种制剂，即加利福尼亚的泥浆章鱼O. bimaculoides。迄今为止，他们已经开发了用于外OVO培养的方法，并为从胚胚到孵化的胚胎阶段进行了整个植物的原位杂交方法。研究人员还分离了一大批cDNA，用于关键发育控制基因。为了完成此模型系统，他们需要在这些头足类胚胎中开发可靠的基因操纵方法。具体目标1是生成基于章鱼EF1-A和泛素C基因的头足类型表达载体。具体目的2是开发向章鱼胚胎递送的质粒方法，研究人员将使用阳离子聚合物作为方法和化学转基因测试。基因操纵资源和方法的开发将允许对头足动物发育的分子和细胞机制进行假设驱动的研究。