Tools for gene editing in marmosets

狨猴基因编辑工具

• 批准号: 10818971
• 负责人: Richard L Huganir
• 金额: $ 16.37万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 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 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10818971
• 关键词: Assisted Reproductive Technology; Brain; Brain Diseases; Breeding; CRISPR/Cas technology; Callithrix; Caring; Cre driver; Development; Embryo; Future; Generations; Genes; Genetic; Genetic Models; Goals; Human; In Vitro; Infrastructure; Methods; Modeling; Mutation; Neurobiology; Ovarian Stimulations; Physiology; Techniques; Technology; Tissues; behavior test; cell type; embryo culture; experimental study; genetic manipulation; genome editing; improved; neuropsychiatry; novel; oocyte retrieval; preimplantation; promoter; screening; selective expression; tool

Tools for Gene Editing in Marmosets SUMMARY The goal of this proposal is to create an efficient new set of tools and techniques for the generation of genetically modified marmosets suitable for use as models of brain physiology and function. Development of efficient methods of genetic manipulation in the species will require refinement of genome editing technologies, breeding technologies, ovarian stimulation and oocyte retrieval, embryo culture, and pre-implantation screening of embryos for the desired genetic alternations. A large colony of marmosets will be necessary, along with an infrastructure to refine technologies and an interdisciplinary team with expertise in marmoset care, assisted reproduction technologies, tissue-specific expression with Cre-drivers and CRISPR technologies, and behavior testing and neuropsychiatric analysis. The Specific Aims of this proposal are to: (1) Improve technology for in vitro genetic manipulation of marmosets and explore novel methods for marmoset gene editing; (2) Identify safe harbor loci and establish Cas9, Cre, and GCaMP marmoset lines using promoters for ubiquitous expression, selective expression in brain, or regional or cell-type specific expression; and (3) Generate marmoset models with genetic alterations relevant to the neurobiology of human brain diseases. These studies will provide tools for future gene editing experiments in marmosets, will establish an initial set of genetically modified tool lines for use in the field, and will provide initial proof-of-principle lines for marmoset models of genetic alterations potentially relevant to the neurobiology of human brain diseases.

狨猴基因编辑工具 概括 该提案的目标是创建一套高效的新工具和技术来生成 转基因狨猴适合用作大脑生理学和功能的模型。发展 物种基因操纵的有效方法将需要改进基因组编辑技术， 育种技术、卵巢刺激和卵母细胞取出、胚胎培养和植入前筛查 胚胎进行所需的基因改变。需要一大群狨猴，以及一只 完善技术的基础设施和具有狨猴护理专业知识的跨学科团队，协助 生殖技术、使用 Cre 驱动程序和 CRISPR 技术的组织特异性表达以及行为 测试和神经精神分析。该提案的具体目标是： (1) 改进in技术 狨猴的体外遗传操作，探索狨猴基因编辑的新方法； (2) 识别安全 拥有基因座并使用启动子建立 Cas9、Cre 和 GCaMP 狨猴品系以实现普遍表达， 大脑中选择性表达，或区域或细胞类型特异性表达； (3) 生成狨猴模型 与人类大脑疾病的神经生物学相关的基因改变。这些研究将提供工具 对于未来的狨猴基因编辑实验，将建立一套初始的转基因工具系 在该领域使用，并将为狨猴基因改变模型提供初步的原理证明 与人类大脑疾病的神经生物学潜在相关。