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

开发海胆胚胎稳定遗传修饰的基础构件

• 批准号: 10575685
• 负责人: AMRO M HAMDOUN
• 金额: $ 23.7万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-12-15 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10575685
• 关键词: Acceleration; Adult; Affect; Animal Genetics; Animal Model; Binding; Biogenesis; Biological; Biomedical Research; CRISPR/Cas technology; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Communities; Cre-LoxP; Cyclins; DNA; DNA Transposable Elements; Development; Disease; Embryo; Fertility; Fertilization; Foundations; Gene Expression Regulation; Gene Transfer Techniques; Generations; Genes; Genetic; Genetic Recombination; Genome; Genome engineering; Goals; Health; Human; Injections; Integrase; Investments; Knock-out; Knowledge; Laboratories; Life; Lytechinus; Mediating; Membrane; Messenger RNA; Methodology; Methods; Mitotic; Modeling; Modernization; Modification; Mutagenesis; National Institute of Child Health and Human Development; National Institute of Environmental Health Sciences; National Institute of General Medical Sciences; Nobel Prize; Nonhomologous DNA End Joining; Optics; Organism; Outcome; Phenotype; Phylogenetic Analysis; Poison; Production; Reporter; Reproducibility; Reproduction; Research; Research Infrastructure; Research Personnel; Resource Sharing; Resources; Sea Urchins; Seminal; Site; Specificity; Structure; Study models; Techniques; Technology; Time; Tissues; Toxic Environmental Substances; Toxic effect; Transgenic Organisms; Transmembrane Transport; Transposase; United States National Institutes of Health; Validation; Work; animal resource; combinatorial; design; early life exposure; experimental study; fluorescence imaging; gene regulatory network; genetic approach; genetic manipulation; human disease; improved; innovation; malformation; model organism; mutant; pathogen; programs; promoter; recombinase-mediated cassette exchange; repaired; success; tool; transgene expression

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.

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。