Enhancing the C. elegans animal resource through genome editing

通过基因组编辑增强线虫动物资源

• 批准号: 10453663
• 负责人: Ann E. Rougvie
• 金额: $ 60.52万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10453663
• 关键词: ATP phosphohydrolase; Address; Alleles; Animal Model; Animals; Area; Attention; Behavior; Biology; Biomedical Research; CRISPR/Cas technology; Caenorhabditis; Caenorhabditis elegans; Candidate Disease Gene; Categories; Clustered Regularly Interspaced Short Palindromic Repeats; Code; Collection; Communities; Databases; Deposition; Development; Disease; Dissection; Equilibrium; Event; Funding; G-Protein-Coupled Receptors; Generations; Genes; Genetic; Genome; Goals; Gold; Guide RNA; Hand; Health; Human; Human Biology; Human Genetics; Information Distribution; Institutes; Invertebrates; Ion Channel; Knock-in; Knock-out; Laboratories; Lead; Light; Methods; Microinjections; Modeling; Molecular; Molecular Biology; Molecular Genetics; Monitor; Mutation; Neurons; Oligonucleotides; Orthologous Gene; Pathway interactions; Peptide Hydrolases; Phase; Phenotype; Phosphotransferases; Physiology; Production; Proteins; Proteomics; Quality Control; Reagent; Reproducibility; Research; Research Personnel; Research Project Grants; Research Support; Resources; Scientist; Site; United States National Institutes of Health; Variant; WormBase; animal resource; base; community center; design; gene discovery; gene function; gene interaction; genetic analysis; genome editing; genome resource; human disease; interest; knockout gene; loss of function; loss of function mutation; metabolomics; mutant; programs; screening; transcriptome sequencing; trend; variant of unknown significance; web site

Project Summary/Abstract The objective of this project is to extend the C. elegans genetic toolkit available to researchers by providing publicly available gene knockout (KO) mutations in genes of high value to those interested in human biology and disease. C. elegans is a proven model for discovery of gene function and for embedding genes into functional pathways, many of which were discovered in this transparent animal and are conserved in humans. Loss-of-function mutations are the gold-standard for genetic analysis and allow inferences of gene function and interaction. A relatively recent trend is that large-scale screens (e.g., RNA-seq, proteomics, natural variation studies) generate long candidate gene lists for researchers to sort through and functionally validate, requiring loss of function mutants in many genes to be examined. Having a community-driven resource generate KOs in such genes provides reproducibility and an economy of scale that benefits all. We have developed an efficient and high throughput CRISPR-Cas9 based gene knockout (KO) project to provide a set of precisely edited gene knockout strains to the communities of C. elegans researchers and human geneticists. We seek to continue the production phase of this project, thereby transformatively increasing the efficiency and impact of C. elegans molecular genetics. We employ a highly coordinated three-site production strategy to generate KOs. Each gene edit is carefully confirmed, and validated strains are grossly phenotyped and deposited into the Caenorhabditis Genetics Center (CGC) strain collection along with detailed strain information for distribution to the community. Strains are advertised through both the CGC and WormBase websites. We have generated over 1,000 KOs to date; these KOs have supported a variety of research projects – spanning from mechanistic studies of human variants to metabolomics – funded by at least 14 NIH Institutes and Centers, demonstrating the high impact of our resource. We propose to continue to use our established pipelines to target an additional 2500 C. elegans orthologs of human genes. We will prioritize known or suspected human disease genes, druggable gene classes, as well as understudied genes that are conserved to humans but that have no actionable information. This is a multi-PI project which includes the lead-PIs of the CGC and of WormBase and a subcontractor who maintains multiple community-centered databases, including a CRISPR guide RNA selection site.

项目摘要/摘要 该项目的目的是通过提供研究人员来扩展可用于研究人员的C. C. 对人类生物学感兴趣的人具有高价值基因的公开基因敲除（KO）突变 和疾病。秀丽隐杆线虫是发现基因功能和将基因嵌入基因的验证模型 功能途径，其中许多是在这种透明动物中发现的，并且在人类中是保守的。 功能丧失突变是遗传分析的金标准，并允许推断基因功能和 相互作用。一个相对较新的趋势是大规模屏幕（例如RNA-Seq，蛋白质组学，自然变化 研究）生成长期候选基因列表，以供研究人员进行分类和验证，需要 许多基因的功能突变体丧失。拥有社区驱动的资源会产生KOS 这样的基因提供了可重复性和有益于所有人的规模经济。我们已经开发了一个高效的 和高吞吐量CRISPR-CAS9基因敲除（KO）项目，以提供一组精确编辑的基因 淘汰赛果岭研究人员和人类遗传学家的社区。我们寻求继续 该项目的生产阶段，从而改变了秀丽隐杆线虫的效率和影响 分子遗传学。我们采用高度协调的三站生产策略来生成KOS。每个 仔细确认基因编辑，并严重表现出经过验证的菌株并沉积在 Caenorhabditis Genetics Center（CGC）菌株收集以及详细的应变信息以分配到 社区。菌株正在通过CGC和Wormbase网站进行广告。我们已经生成了 迄今为止，超过1,000个KO；这些KO支持了各种研究项目 - 跨越机械 对代谢组学的人类变体的研究 - 由至少14个NIH机构和中心资助，证明 我们资源的高影响。我们建议继续使用我们既定的管道来瞄准 2500 C.人类基因的秀丽隐杆线虫。我们将优先考虑已知或怀疑的人类疾病基因， 可吸毒基因类别，以及对人类保守但没有的理解基因 可行的信息。这是一个多P-PI项目，包括CGC和Wormbase和Wormbase和 分包商维护多个以社区为中心的数据库，包括CRISPR指南RNA 选择网站。