Tools for gene editing in marmosets

狨猴基因编辑工具

基本信息

批准号：
10508541
负责人：
Richard L Huganir
金额：
$ 179.82万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
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/10508541
关键词：
3-Dimensional Assisted Reproductive Technology Behavior Behavioral Brain Brain Diseases Breeding CRISPR/Cas technology CUL1 gene Callithrix Callithrix jacchus jacchus Caring Cognitive Communication Research Communities Cre driver DNA cassette Development Electroporation Embryo Embryo Transfer Fertilization in Vitro Freezing Functional disorder Future Gene Expression Generations Genes Genetic Genetic Models Germ Cells Goals Human In Vitro Infrastructure Methods Modeling Modification Molecular Mutation Neurobiology Neurons Neurosciences Ovarian Stimulations Penetrance Phenotype Physiology Population Pregnancy Preimplantation Diagnosis Preparation Procedures Protocols documentation Reporter Research Rodent SYNGAP1 Spermatids Sterility Syndrome System Techniques Technology Teratoma Testing Testis Tissues Universities behavior test brain cell cell type cognitive function cost embryo culture experience experimental study frontal lobe genetic manipulation genome editing genomic locus improved in vivo in vivo Model male neurobiological mechanism neuropsychiatric disorder neuropsychiatry nonhuman primate novel oocyte maturation oocyte retrieval preimplantation promoter screening selective expression social social communication sperm cell sperm viability stem cells tool

项目摘要

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。