Rat Germline Gene Editing Products and Services

摘要 捐赠者精原干细胞的直接种系编辑将简化遗传生产 改良的模型生物，并提供前所未有的质量控制，以准确预测生殖系 单独选择或多个共同选择的目标等位基因的传播率。 GenomeDesign的直接生殖系编辑技术将通过以下方式改变定制大鼠模型市场 在最广泛的范围内提供最广泛的目标基因组DNA插入大小 易驯服的老鼠遗传背景。在这里，我们提出了生产冷冻产品的三个具体目标 精原细胞库可用作生殖系载体，以构建定制的遗传设计 改良大鼠模型。在具体目标1中，精原细胞来源于多达12种不同的流行 将产生近交系和近交系大鼠的遗传背景，以提供最多的基因组设计 全面的目录定制可设计的老鼠模型。在特定目标2中，精准精原 基因打靶效率将通过滴状数字聚合酶链式反应(DdPCR)来测量，以指导动物 生产协议，使个人客户和基因设计公司受益。我们将把ddPCR应用于 通过共传播、共选择的基因打靶获得的项目实际生产成本和时间节省 来自单个生殖系的事件，或来自不同精原系的多克隆大鼠生产的事件 被汇集并移植到一只受体大鼠体内。在具体目标3中，发展受援国男性 与不同的捐赠者精原菌株兼容将建立一个无与伦比的生物医学组合 适用于基因工程的相关大鼠品系。通过完成第一阶段的特定目标， GenomeDesign将提供概念验证，以建立具有遗传多样性的 精原种子库，使精确的、有性的、强有力的遗传选择成为可能 直接在大鼠生殖系中进行可传递的基因组修改。 这一SBIR第一阶段项目的基准是： 1)将精原基因打靶服务显著扩展到新的大鼠遗传背景 2)建立ddpcr作为共同选择的基因靶向事件的生殖系传播的预测 3)建立不同供体大鼠精原遗传学背景下的受体大鼠模型

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