Rat Germline Gene Editing Products and Services

大鼠种系基因编辑产品和服务

• 批准号: 9409628
• 负责人: F. Kent Hamra
• 金额: $ 17万
• 依托单位: GENOMEDESIGNS LABORATORY, LLC
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9409628
• 关键词: Adult; Alleles; Allogenic; Animal Model; Animals; Benchmarking; Biological Assay; Biological Process; Biomedical Research; Businesses; Catalogs; Cell Culture Techniques; Custom; Event; Fertility; Freezing; Gene Targeting; Genes; Genetic; Genetic Engineering; Genetic Models; Genomic DNA; Genomics; Health; Human; Inbreeding; Individual; Measures; Methods; Modeling; Modification; Organism; Phase; Production; Protocols documentation; Quality Control; Rat Strains; Rattus; Research Personnel; Savings; Scientist; Seeds; Services; Small Business Innovation Research Grant; Stem cells; Technology; Testis; Time; Transplantation; clinically relevant; cost; design; digital; gene function; genetic selection; germline stem cells; male; mutant; novel; prognostic; sperm cell; transmission process; vector

Summary Direct germline editing in donor spermatogonial stem cells will streamline production of genetically modified model organisms and provide unprecedented quality control to accurately predict germline transmission rates for individually selected, or multiple, co-selected targeted alleles. GenomeDesigns' direct germline editing technologies will transform the custom rat model market by providing the broadest range of targeted genomic DNA insertion sizes available in the broadest range of tractable rat genetic backgrounds. Here, we propose three specific aims to produce frozen spermatogonial stocks that can be used as germline vectors to build custom designed, genetically modified rat models. In Specific Aim 1, spermatogonial stocks derived from up to 12 different popular outbred and inbred rat genetic backgrounds will be generated to seed GenomeDesigns with the most comprehensive catalogue of custom designable rat models. In Specific Aim 2, precise spermatogonial gene targeting efficiencies will be measured by droplet digital PCR (ddPCR) to guide animal production protocols, benefiting individual customers and GenomeDesigns. We will apply ddPCR to project real production cost and time savings received by co-transmitting, co-selected gene targeting events from an individual germline, or by polyclonal rat production where distinct spermatogonial lines are pooled and transplanted into a single recipient rat. In Specific Aim 3, developing recipient males compatible with diverse donor spermatogonial strains will build an unmatched portfolio of biomedically relevant rat strains amenable to genetic engineering. By completing Phase I Specific Aims, GenomeDesigns will provide proof-of-concept for building a genetically diverse catalogue of spermatogonial stocks that uniquely enable strong genetic selection for precise, sexually transmittable, genomic modifications directly in the rat germline. Benchmarks for this Phase I SBIR project are: 1) Significantly expand spermatogonial gene targeting services into new rat genetic backgrounds 2) Establish ddPCR as prognostic for germline transmission of co-selected gene targeting events 3) Produce recipient rat models for diverse donor rat spermatogonial genetic backgrounds

概括 供体精原干细胞的直接种系编辑将简化遗传物质的生产 修改模型生物并提供前所未有的质量控制以准确预测种系 单独选择的或多个共同选择的目标等位基因的传输率。 GenomeDesigns 的直接种系编辑技术将通过以下方式改变定制大鼠模型市场： 提供最广泛的目标基因组 DNA 插入尺寸 易驯服的大鼠遗传背景。在这里，我们提出了生产冷冻食品的三个具体目标 精原细胞库存可用作种系载体来构建定制设计的遗传基因 改良的大鼠模型。在具体目标 1 中，精原细胞来源于多达 12 种不同的流行 将生成远交和近交大鼠遗传背景，以种子基因组设计具有最多 定制可设计大鼠模型的综合目录。在具体目标 2 中，精确精原细胞 基因靶向效率将通过液滴数字 PCR (ddPCR) 来测量，以指导动物 生产协议，使个人客户和 GenomeDesigns 受益。我们将应用 ddPCR 来 项目通过共同传输、共同选择的基因靶向所节省的实际生产成本和时间 来自单个种系的事件，或多克隆大鼠的生产，其中不同的精原细胞系 汇集并移植到单个受体大鼠体内。在具体目标 3 中，培养受体男性 与不同供体精原细胞菌株兼容将建立一个无与伦比的生物医学组合 适合基因工程的相关大鼠品系。通过完成第一阶段的具体目标， GenomeDesigns 将为构建基因多样性目录提供概念验证 精原细胞种群具有独特的能力，可以对精确的性行为进行强有力的遗传选择 直接在大鼠种系中进行可传递的基因组修饰。 第一阶段 SBIR 项目的基准是： 1) 将精原基因靶向服务显着扩展到新的大鼠遗传背景 2) 建立 ddPCR 作为共选基因靶向事件种系传播的预后 3) 制作不同供体大鼠精原遗传背景的受体大鼠模型