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.

项目概要/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 以及更好地了解动物生物学。动物基因组的定向修饰使得 动物呈现人类疾病表型，因此，对于设计和开发新的治疗方法至关重要。 使用猪等 GE 大型动物模型通常会产生与临床相关的结果，因为它们的生理学 和解剖学类似于人类。例如，引入导致囊性纤维化的遗传元件和 猪基因组的免疫缺陷导致转基因猪密切再现疾病的症状。然而， 由于建立此类模型需要大量时间，转基因猪在生物医学中尚未广泛应用。作为 对于大型动物品种，猪的一轮繁殖至少需要一年的时间，而且通常需要多轮 育种对于建立转基因猪模型是必要的。 CRISPR/Cas等基因组编辑工具的应用 系统，显着提高了对猪基因组进行靶向修饰的效率。然而， 对基因组编辑程序造成的意外基因组改变以及引入所需时间的担忧 对猪作为大型动物模型的针对性改造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的有效性和安全性并设计新方法 这将有助于在有针对性的基因组编辑事件后对动物模型进行快速表型分析。三个具体目标 旨在实现我们的目标。首先，我们将生成用于全局检测 GE 脱靶事件的方法 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， 我们建议开发一种策略来快速对 GE 胎儿进行表型分析并修改野生型猪的基因组。 第三个目标的目标是与农业和生物医学相关性状相关的基因。 从该项目中获得的知识可用于扩大转基因猪在生物医学中的应用，同时 对农业生产也产生影响。使用基因组预测表型的重要性 为了快速鉴定改良的等位基因和性状，将在这里种植。我们在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。该项目的成果应 通过有效捕获后续信息，提高基因工程猪模型在生物医学和农业中的可用性 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该药物的功效 拟议的战略；然而，我们的研究结果应该很容易转移到生产其他动物模型 生物医学和农业上重要的物种也是如此。鉴于猪用作动物模型的重要性， 我们的研究结果应该对美国国立卫生研究院和美国农业部机构都有好处。