Development of foundational building blocks for stable genetic modification of sea urchin embryos

项目摘要 动物模型对于理解发育机制和 揭示疾病的早期起源这些模型的效用取决于有效的 应用遗传学方法建立基因和表型之间的因果关系。 该提案的目标是针对研究基础设施计划办公室（ORIP）， 建立资源，大大提高研究的可重复性，实用性和效率 在海胆-这是由数百名研究人员在众多国家卫生研究所支持使用- 项目这些资源包括模块化的创始人线路（“构建块”）， 方法，从细胞和阶段特异性诱变到组合荧光报告基因 表达，到活跃的遗传学。该提案的目的是利用传统的（转座子- 介导的转基因）和现代（CRISPR/Cas9）技术来满足这些不同的需求。 这项工作的进一步创新来自于将这种模块化遗传设计应用于一个物种 具有理想的生物学特征和转基因和研究的知识背景- 规模化培养这项工作将对跨任务的研究产生变革性影响， 包括生殖、膜转运和基因调控的研究。

PROJECT SUMMARY Animal models are essential for understanding the mechanisms of development, and for uncovering the early life origins of disease. The utility of these models depends on the effective application of genetic methods to establish causal relationships between genes and phenotypes. The goal of this proposal, targeted to the Office of Research Infrastructure Programs (ORIP), is to build resources that dramatically improve the reproducibility, utility, and efficiency of research in sea urchins - which are used by hundreds of researchers across numerous NIH-supported projects. These resources include modular founder lines (“building blocks”) enabling myriad approaches, from cell and stage-specific mutagenesis to combinatorial fluorescent reporter expression, to active genetics. The Aims of this proposal utilize both traditional (transposon- mediated transgenesis) and modern (CRISPR/Cas9) techniques to meet these diverse needs. Further innovation of the work comes from application of this modular genetic design to a species with the ideal biological features and background of knowledge for transgenesis and research- scale culturing. This work will have transformative impact on research across the mandates of NIH, including studies on reproduction, membrane transport, and gene regulation.