Enhancing The Value of Pigs for Agriculture and Biomedical Applications By Using Novel Genome Editing Strategies

通过使用新型基因组编辑策略提高猪在农业和生物医学应用中的价值

• 批准号: 10755034
• 负责人: Kiho Lee
• 金额: $ 78.18万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10755034
• 关键词: Agriculture; Alleles; Anatomy; Animal Model; Animals; Area; Biochemical; Biological; Biological Assay; Biology; Breeding; CRISPR/Cas technology; Clinic; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Cystic Fibrosis; Data; Detection; Disease; Early Diagnosis; Embryo Cloning; Event; Exclusion; Family suidae; Fetus; Food production; Genes; Genetic; Genetic Diseases; Genetic Engineering; Genome; Genomics; Goals; Heritability; Human; Immune; Immune response; Immunologic Deficiency Syndromes; Investments; Knowledge; Length; Life Cycle Stages; Link; Livestock; Methods; Modeling; Modification; Molecular; Monitor; Outcome; Patients; Phenotype; Physiology; Porcine respiratory and reproductive syndrome virus; Pre-Clinical Model; Pregnancy; Procedures; Process; Production; Puberty; Research; Research Activity; Resistance; Resources; Safety; Secure; Site-Directed Mutagenesis; Somatic Cell; Symptoms; System; Technology; Testing; Time; United States Department of Agriculture; United States National Institutes of Health; Variant; clinical effect; clinical outcome assessment; clinical phenotype; clinically relevant; design; detection platform; disease phenotype; efficacy evaluation; fetal; gene function; gene therapy; genetic element; genome editing; genome integrity; genome sequencing; human disease; improved; in vivo; inhibitor; novel; novel strategies; pig genome; porcine model; postnatal; predictive tools; preservation; response; side effect; single-cell RNA sequencing; small molecule; therapy outcome; tool; trait; whole genome

Project Summary/Abstract Genetically engineered (GE) animal models are essential for generating biomedical models for human disease and for gaining a better understanding of animal biology. Targeted modification of the animal genome allows the animals to present human disease phenotypes, and therefore, are critical to design and develop novel treatments. The use of GE large animal models such as pigs often results in clinically relevant outcomes as their physiology and anatomy resemble humans. For example, introducing genetic elements responsible for cystic fibrosis and immunodeficiency to the pig genome induces GE pigs to closely recapitulate symptoms of the diseases. However, GE pigs are not widely available in biomedicine due to the amount of time required to establish such models. As a large animal species, a single round of breeding in pigs takes at least one year and often multiple rounds of breeding is necessary to establish GE pig models. Application of genome editing tools, such as the CRISPR/Cas system, has significantly improved efficacy to introduce targeted modifications to the pig genome. However, concerns over unintended genome alterations from genome editing procedure and days required to introduce targeted modifications in pigs as a large animal model impedes wide use of the technology. Our objective of this project is to evaluate the efficacy and safety of genome editing technology and design novel approaches that will assist in rapid phenotyping of animal models after a targeted genome editing event. Three specific aims are proposed to reach our goal. First, we will generate methods for global detection of off-targeting events in GE pigs. Secondly, we will develop strategies to secure genome integrity during the genome editing process. Finally, we propose to develop a strategy to rapidly phenotype GE fetuses and to modify the genome of wild-type pigs. Targets of this third aim are genes associated with traits that are relevant in both agriculture and biomedicine. The knowledge obtained from this project can be implemented to expand the use of GE pigs in biomedicine while also having an impact on agriculture production. The importance of using the genome to predict the phenotype for rapid identification of improved alleles and traits will be grown here. Our expertise in using genome editing technology and GE pig models will be employed to complete the proposed aims. Outcomes of this project should increase the availability of GE pig models in biomedicine and agriculture by effectively capturing subsequent phenotypes after genome editing events. We propose to utilize pigs as a model to investigate the efficacy of the proposed strategies; however, our findings should be easily transferred to producing other animal models in biomedicine and agriculturally important species, as well. Given the importance of pigs used as animal models, our findings should be beneficial to both NIH and USDA agencies.

项目摘要/摘要 基因工程（GE）动物模型对于生成人类疾病的生物医学模型至关重要 并获得对动物生物学的更好理解。动物基因组的靶向修饰允许 动物呈现人类疾病表型，因此对于设计和开发新疗法至关重要。 使用GE大动物模型（例如猪）通常会导致临床相关结果作为其生理学 解剖学类似于人类。例如，引入负责囊性纤维化和 对猪基因组的免疫缺陷会诱导GE猪密切概括疾病的症状。然而， 由于建立此类模型所需的时间，GE猪在生物医学中并未广泛使用。作为 一种大动物物种，一轮猪的繁殖至少需要一年，通常需要多个回合 建立GE猪模型是必要的。基因组编辑工具的应用，例如CRISPR/CAS 系统，具有显着提高的功效，以将靶向修饰引入猪基因组。然而， 对基因组编辑程序的意外基因组改变的担忧以及引入所需的天数 猪作为大型动物模型的靶向修改阻碍了该技术的广泛使用。我们的目标 该项目旨在评估基因组编辑技术和设计新颖方法的功效和安全性 这将有助于在靶向基因组编辑事件后快速表型对动物模型进行快速分型。三个具体目标 建议实现我们的目标。首先，我们将生成用于全局检测GE中靶向事件的方法 猪。其次，我们将制定在基因组编辑过程中确保基因组完整性的策略。最后， 我们建议制定一种策略来快速表型GE胎儿并修改野生型猪的基因组。 这第三个目标的目标是与农业和生物医学既相关的性状相关的基因。 可以实施从该项目中获得的知识，以扩大生物医学中的Ge Pigs的使用 也对农业生产产生影响。使用基因组预测表型的重要性 为了快速鉴定改善的等位基因和特征。我们使用基因组编辑方面的专业知识 将采用技术和GE猪模型来完成拟议的目标。这个项目的结果应该 通过有效捕获随后的生物医学和农业中GE Pig模型的可用性 基因组编辑事件后的表型。我们建议利用猪作为模型来研究 拟议的策略；但是，我们的发现应轻松转移到生产其他动物模型中 生物医学和农业重要物种也是如此。考虑到用作动物模型的猪的重要性， 我们的发现应该对NIH和USDA机构都有益。