High-throughput gene disruption in zebrafish using retroviral integration

描述（申请人提供）：斑马鱼已被广泛用作脊椎动物发育和遗传学研究的模型动物。众所周知，对斑马鱼基因的遗传研究可以深入了解保守脊椎动物基因的功能，从而更好地了解人类疾病和健康。随着斑马鱼基因组测序计划的接近完成，我们非常希望通过测序预测的所有基因也将携带一个易于用于基因功能研究的突变。为了实现这一目标，我们开发了一个高效的简化平台，使用前病毒插入加上高通量测序和定位技术来诱变斑马鱼基因组中基本上所有可识别的候选基因。在这个项目中，我们建议通过构建一个斑马鱼菌株文库，为公共研究界提供遗传工具，其中大约有12,000个斑马鱼基因被逆转录病毒插入破坏。所有生成的资源将在出版前以最低的成本提供给社区。公共卫生相关性：斑马鱼是一种受欢迎的脊椎动物模型生物，用于研究发育和人类疾病，我们使用最先进的技术在大多数斑马鱼基因中产生突变。由于这些基因高度保守，我们的研究将为理解人类疾病和协助设计新的治疗方法提供有价值的信息。

DESCRIPTION (provided by applicant): Zebrafish has been widely used as a model animal to study vertebrate development and genetics. It is well established that genetic studies of zebrafish genes can provide insights into the function of the conserved vertebrate genes and therefore lead to better understanding of human diseases and health. With the zebrafish genome sequencing project nearly completed, it is highly desirable that all of the genes predicated by sequencing will also carry a mutation that readily available for gene function studies. Towards this goal, we have developed an efficient and streamlined platform using proviral insertions coupled with high-throughput sequencing and mapping technologies to mutagenize essentially all of the identifiable candidate genes in zebrafish genome. In this project we propose to provide genetic tools to the public research community by constructing a library of zebrafish strains in which approximately 12,000 zebrafish genes have been disrupted with a retrovirus insertion. All the resources generated will be made available to the community with a minimal cost before publication. PUBLIC HEALTH RELEVANCE: Zebrafish is a popular vertebrate model organism for studying development and human diseases and we use state of the art technologies to generate mutations in most of zebrafish genes. Since these genes are highly conserved our studies will provide valuable information for understanding human diseases and assisting design of novel therapeutic approaches.