A Genetic Engineering Toolbox for Marmosets (GETMarm): Development and optimization of genome editing and assisted reproduction techniques for marmoset models

狨猴基因工程工具箱 (GETMarm)：狨猴模型基因组编辑和辅助生殖技术的开发和优化

• 批准号: 10459550
• 负责人: Guoping Feng
• 金额: $ 112.04万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10459550
• 关键词: Alleles; Animal Model; Animals; Area; Assisted Reproductive Technology; Biology; Biopsy Specimen; Brain; Brain Diseases; Breeding; Callithrix; Callithrix jacchus jacchus; Cell model; Chromosomes; Clustered Regularly Interspaced Short Palindromic Repeats; Cryopreservation; Deposition; Development; Diagnostic; Disease model; Embryo; Embryo Transfer; Engineering; Enhancers; Essential Genes; Evaluation; Female; Frequencies; Functional disorder; Gene Frequency; Generations; Genetic; Genetic Engineering; Genetic Models; Genome; Genome engineering; Genotype; Goals; Human; Injections; Knock-in; Macaca; Methods; Modeling; Modernization; Molecular; Mosaicism; Mus; Neurosciences; Neurosciences Research; Oocytes; Persons; Preimplantation Diagnosis; Primates; Process; Protocols documentation; Reagent; Reproductive Techniques; Research; Resource Sharing; Rett Syndrome; Rodent; Rodent Model; Sexual Maturation; Single Nucleotide Polymorphism; Standardization; Structure; Study models; Synaptic plasticity; System; Technology; Testing; Time; Training; Transposase; United States National Institutes of Health; Work; assisted reproduction; autism spectrum disorder; base; base editing; base editor; cost; data hub; embryo culture; genetic manipulation; genetic testing; genome editing; improved; in vivo; infancy; mutant; nonhuman primate; novel; oocyte maturation; prenatal; prime editing; programs; repaired; tool; trait; translational neuroscience; zygote

PROJECT SUMMARY While mice are essential models for many areas of neuroscience, there are also many aspects of higher brain function and dysfunction that cannot be adequately modeled in rodents. Thus, there is a need for new genetic models that have brain structure and function closer to humans. For these reasons, non-human primates (NHP) provide an attractive model to study higher brain function and brain disorders. A promising emerging NHP model is the common marmoset, a small New World primate that has many advantages as a genetic model. Although the adaptation of bacterial CRISPR/Cas systems for targeted genome engineering and model creation has revolutionized modern biology, editing of the marmoset genome is still in its infancy. Given the significant time and money required for marmoset genome editing, new methods to increase editing efficiency, decrease mosaicism, and identify correctly-edited embryos prior to transfer to recipient females are critical. Additionally, new methods for controlling the zygosity of founder animals are necessary to enable analysis of homozygous F0 animals and to avoid homozygous editing when targeting essential genes that cause embryonic lethality upon biallelic disruption. To these ends, we propose a research program that will significantly enhance our ability to introduce multiple types of edits into the marmoset genome, reduce mosaicism, control the zygosity of edits, and identify successfully edited embryos through prenatal genetic testing. We will disseminate these technologies and models through direct resource sharing, in-person trainings, deposition to NIH-supported Marmoset Coordination Center. Together, the proposed advances will significantly reduce the time, effort, costs and animal numbers necessary to marmoset genetic models and will unlock the true potential of marmosets for basic and translational neuroscience research.

项目摘要 虽然小鼠是神经科学许多领域的重要模型，但也有许多方面 无法在啮齿动物中充分建模的较高的大脑功能和功能障碍。因此，需要 具有大脑结构和功能更接近人类的新遗传模型。由于这些原因，非人类 灵长类动物（NHP）提供了一个有吸引力的模型来研究更高的大脑功能和脑部疾病。有希望的 新兴的NHP模型是普通的Marmoset，这是一种新世界的灵长类动物，具有许多优势 遗传模型。虽然针对靶向基因组工程的细菌CRISPR/CAS系统的适应 模型的创建彻底改变了现代生物学，对Marmoset基因组的编辑仍处于起步阶段。 鉴于Marmoset基因组编辑所需的大量时间和金钱，因此增加了编辑的新方法 在转移到受体女性之前 批判的。此外，必须采用控制创始人动物的合格性的新方法才能实现 分析纯合F0动物，并在针对基本基因时避免纯合编辑 在双重破坏时引起胚胎致死性。对于这些目的，我们提出了一项研究计划，将 显着增强了我们将多种类型的编辑引入Marmoset基因组的能力，减少 镶嵌，控制编辑的糊状，并通过产前遗传确定成功编辑的胚胎 测试。我们将通过直接的资源共享来传播这些技术和模型 培训，向NIH支持的Marmoset协调中心沉积。拟议的进步将在一起 大大减少摩尔马斯遗传模型所需的时间，努力，成本和动物数量，并将 释放了摩尔摩斯群岛对基本和转化神经科学研究的真正潜力。