Rat Germline Gene Editing Products and Services

总结 供体精原干细胞中的直接生殖系编辑将简化遗传学上的生产。 改良的模式生物，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靶向等位基因的传输率。 GenomeDesigns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： 提供最宽范围的靶向基因组DNA插入大小， 温顺的老鼠遗传背景在这里，我们提出了三个具体目标，以生产冷冻 精原细胞储备，可以用作生殖系载体，以建立定制设计的，遗传 改良大鼠模型。在特定目标1中，精原细胞来源于多达12种不同的流行 将产生远系繁殖和近交大鼠遗传背景，以播种具有最多遗传背景的基因组设计。 定制可设计大鼠模型的全面目录。在特定目标2中，精原细胞 将通过液滴数字PCR（ddPCR）测量基因靶向效率，以指导动物 生产协议，使个人客户和GenomeDesigns受益。我们将ddPCR应用于 通过共同传播、共同选择的基因靶向，节省项目真实的生产成本和时间 事件，或通过多克隆大鼠生产，其中不同的精原细胞系 合并并移植到单个受体大鼠中。在具体目标3中，发育中的受体男性 与不同的供体精原细胞株相容，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将为建立一个遗传多样性的基因目录提供概念验证。 精原种群，独特地使强大的遗传选择， 直接在大鼠生殖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)将精原细胞基因靶向服务显著扩展到新的大鼠遗传背景 2)建立ddPCR作为共选择基因靶向事件生殖系传播的预后 3)建立不同供鼠精原细胞遗传背景的受体大鼠模型

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