Genetic Engineering Core

基因工程核心

• 批准号: 10494772
• 负责人: Gregg E. Homanics
• 金额: $ 35.15万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10494772
• 关键词: Abbreviations; Age-Months; Alzheimer' s Disease; Alzheimer' s disease model; Animal Genetics; Animal Model; Animals; Assisted Reproductive Technology; Astrocytes; Biomedical Research; Birth; Brain; Brain Diseases; CRISPR/Cas technology; Callithrix; Callithrix jacchus jacchus; Cell Differentiation process; Cell Line; Clustered Regularly Interspaced Short Palindromic Repeats; Communities; DNA sequencing; Development; Early Onset Alzheimer Disease; Electroporation; Embryo; Embryo Transfer; Fibroblasts; Follicle Stimulating Hormone; Generations; Genes; Genetic Engineering; Genome; Genotype; Goals; Gonadotropins; Human; Human Chorionic Gonadotropin; In Vitro; Infant; Knock-in; Knock-out; Late Onset Alzheimer Disease; Libraries; Methodology; Methods; Microinjections; Modeling; Modification; Mosaicism; Mus; Neurobiology; Neurons; Neurosciences; Oocytes; Ovarian Stimulations; PMSG (Gonadotropins); Phenotype; Pregnancy; Production; Reagent; Services; Speed; Surrogate Mothers; Techniques; Technology; Testing; Validation; cost; design; high risk; improved; inhibin; nerve stem cell; nonhuman primate; novel; novel strategies; offspring; risk variant; translational neuroscience; zygote

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.

项目概要 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 通过为 (1) 使用 CRISPR/Cas9 基因编辑对狨猴进行基因改造提供服务， (2) 开发和优化狨猴种系基因编辑的支持技术，以及 (3) 创建成纤维细胞系并将其分化为诱导神经元干细胞和神经元。 普通狨猴 (Callithrix jacchus) 是一种新世界非人类灵长类动物，拥有大量的 在生物医学研究中具有实际优势，并迅速成为备受追捧的模型 神经科学界的有机体。对狨猴的欣赏迅速增长 神经生物学正在与基因编辑技术的同期进步相结合， 使狨猴成为理解基因工程的非人类灵长类动物 大脑功能并研究阿尔茨海默病等大脑疾病。 GEC 将提供 使用 CRISPR/Cas9 基因编辑产生新型基因工程所需的所有服务 狨猴携带早发性和晚发性阿尔茨海默病的人类高风险等位基因。 在创造这些基因编辑动物的同时，GEC 将同时开发和优化 高效种系基因编辑的支持技术。这些技术包括新颖的 卵巢刺激以增强成熟狨猴卵母细胞的产生的方法，以及新的方法 减少基因编辑狨猴镶嵌现象的方法。改进的技术将减少 降低成本并提高基因工程狨猴的生产速度 同时也减少了动物数量。最后，GEC 将生产狨猴成纤维细胞系 并将其分化为诱导神经元干细胞和神经元。细胞系将用于 创建 DNA 测序文库并用于基因编辑的体外表型表征 动物来支持项目目标。