Genetic Engineering Core

项目总结基因工程核心(GEC) 基因工程核心(GEC)的总体目标是支持Marmo-AD的需求 通过向(1)使用CRISPR/CAS9基因编辑的基因工程绒猴提供服务， (2)开发和优化绒猴生殖系基因编辑的使能技术，以及(3) 建立成纤维细胞系并将其分化为诱导神经干细胞和神经元。 常见的绒猴(Callithrix Jacchus)是一种新大陆的非人类灵长类动物，有许多 在生物医学研究中的实用优势，并正在迅速成为备受追捧的模式 神经科学界的有机体。对绒猴的快速增长的欣赏 神经生物学正在与基因编辑技术的同期进步协同作用，以 让绒猴成为基因工程非人类理解灵长类动物的首选 并研究阿尔茨海默氏症等脑部疾病。GEC将提供 使用CRISPR/Cas9基因编辑生产新型基因工程所需的所有服务 携带人类早发和晚发阿尔茨海默病高危等位基因的绒猴。 在创造这些经过基因编辑的动物的同时，GEC将同时开发和优化 实现有效的生殖系基因编辑的技术。这些技术包括新颖的 提高成熟绒猴卵母细胞产量的卵巢刺激方法和新方法 减少基因编辑绒猴嵌合体的方法。改进的技术将减少 生产基因工程绒猴的成本和速度的提高 同时也减少了动物的数量。最后，GEC将培养出绒猴成纤维细胞系 并将其分化为诱导的神经元干细胞和神经元。细胞系将被用来 创建DNA测序文库并用于编辑的基因的体外表型表征 支持项目目标的动物。

PROJECT SUMMARY GENETIC ENGINEERING CORE (GEC) The overall goal of the Genetic Engineering Core (GEC) is to support the needs of MARMO-AD by providing services to (1) genetically engineer marmosets using CRISPR/Cas9 gene editing, (2) develop and optimize enabling technologies for germline gene editing in marmoset, and (3) create fibroblast cell lines and differentiate them into induce neuronal stem cells and neurons. The common marmoset (Callithrix jacchus) is a New World non-human primate with numerous practical advantages in biomedical research and is quickly becoming a much sought-after model organism by the neuroscience community. The rapidly growing appreciation of marmoset neurobiology is synergizing with contemporaneous advances in gene editing techniques to make marmosets the genetically engineered non-human primate of choice for understanding brain function and to study brain disorders such as Alzheimer’s Disease. The GEC will provide all services required to use CRISPR/Cas9 gene editing to produce novel genetically engineered marmosets that harbor human high-risk alleles for early and late onset Alzheimer’s disease. While creating these gene-edited animals, the GEC will simultaneously develop and optimize enabling technologies for efficient germline gene editing. These technologies include novel methods of ovarian stimulation to enhance production of mature marmoset oocytes, and novel approaches to reduce mosaicism in gene edited marmosets. Improved technologies will reduce the cost and increase the speed at which genetically engineered marmosets can be produced while also reducing animal numbers. Lastly, the GEC will produce marmoset fibroblast cell lines and differentiate them into induced neuronal stem cells and neurons. Cell lines will be used to create DNA sequencing libraries and for in vitro phenotypic characterization of gene edited animals in support of the Project goals.