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.

Marmosets中基因编辑的工具 概括 该提案的目的是为生成的有效的新工具和技术创建一个有效的新工具和技术 转基修饰的摩尔果适合用作脑生理和功能的模型。发展 物种中有效的遗传操作方法将需要改进基因组编辑技术， 繁殖技术，卵巢刺激和卵母细胞检索，胚胎培养和植入前筛查 所需遗传交替的胚胎的胚胎。需要大量的果酱殖民地 优化技术的基础架构和具有Marmoset Care专业知识的跨学科团队，协助 繁殖技术，CRE驱动器和CRISPR技术的组织特异性表达以及行为 测试和神经精神分析。该提案的具体目的是：（1）改善技术的技术 体外遗传操纵果冻，并探索新颖的摩尔果基因编辑方法； （2）确定安全 港口基因座并使用启动子来建立CAS9，CRE和GCAMP Marmoset线，以表达无处不在的表达 在大脑或区域或细胞类型的特定表达中的选择性表达； （3）生成马尔莫斯特模型 与人脑疾病的神经生物学有关的遗传改变。这些研究将提供工具 对于Marmoset中的未来基因编辑实验，将建立一组初始的转基因工具线 在现场使用，并将为遗传变化的摩尔马斯群岛模型提供初始的原理证明 可能与人脑疾病的神经生物学有关。