Enhancing the C. elegans animal resource through genome editing

通过基因组编辑增强线虫动物资源

基本信息

批准号：
10610879
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
项目类别：
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/10610879
关键词：
ATP phosphohydrolase Acceleration 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 Event Funding G-Protein-Coupled Receptors Generations Genes Genetic Genome Goals Guide RNA Hand Health Human Human Biology Human Genetics Information Distribution Invertebrates Ion Channel Knock-in Knock-out Laboratories Lead MET gene 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 Sorting United States National Institutes of Health Variant WormBase animal resource community center design gene conservation 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 model organism mutant programs screening transcriptome sequencing trend variant of unknown significance web site

项目摘要

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.

项目概要/摘要该项目的目标是通过提供研究人员可用的线虫遗传工具包对于那些对人类生物学感兴趣的人来说，公开的基因敲除（KO）突变具有很高的价值线虫是发现基因功能和将基因嵌入的经过验证的模型。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黄金标准，可以推断基因功能和一个相对较新的趋势是大规模筛选（例如RNA-seq、蛋白质组学、自然变异）。研究）生成长候选基因列表，供研究人员进行分类和功能验证，需要许多待检测基因的功能缺失突变体具有社区驱动的资源，可在以下基因中产生 KO。这些基因提供了可重复性和规模经济，使所有人受益。以及基于高通量 CRISPR-Cas9 的基因敲除 (KO) 项目，提供一组精确编辑的基因我们寻求继续向秀丽隐杆线虫研究人员和人类遗传学家群体提供敲除菌株。该项目的生产阶段，彻底提高了线虫的效率和影响我们采用高度协调的三位点生产策略来生成 KO。基因编辑经过仔细确认，并对经过验证的菌株进行粗表型分析并存入秀丽隐杆线虫遗传学中心 (CGC) 收集菌株以及详细的菌株信息，以便分发给我们生成的菌株通过 CGC 和 WormBase 网站进行宣传。迄今为止已有 1,000 多个 KO；这些 KO 已支持各种研究项目——从机械学到代谢组学的人类变异研究——由至少 14 个 NIH 研究所和中心资助，证明我们建议继续利用我们现有的渠道来瞄准更多目标。 2500 个人类基因的线虫直系同源物我们将优先考虑已知或可疑的人类疾病基因，可药物基因类别，以及人类保守但没有得到充分研究的基因这是一个多 PI 项目，其中包括 CGC 和 WormBase 的主要 PI 以及维护多个以社区为中心的数据库（包括 CRISPR 引导 RNA）的分包商选择网站。

项目成果

期刊论文数量（12）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Modular metabolite assembly in Caenorhabditis elegans depends on carboxylesterases and formation of lysosome-related organelles.

秀丽隐杆线虫中的模块化代谢物组装取决于羧酸酯酶和溶酶体相关细胞器的形成。

DOI：
发表时间：
2020
期刊：
影响因子：
7.7
作者：
Le, Henry H;Wrobel, Chester Jj;Cohen, Sarah M;Yu, Jingfang;Park, Heenam;Helf, Maximilian J;Curtis, Brian J;Kruempel, Joseph C;Rodrigues, Pedro Reis;Hu, Patrick J;Sternberg, Paul W;Schroeder, Frank C
通讯作者：
Schroeder, Frank C

Identification of essential genes in Caenorhabditis elegans through whole-genome sequencing of legacy mutant collections.

通过遗留突变体集合的全基因组测序鉴定秀丽隐杆线虫的必需基因。