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.

项目总结/摘要 基因工程动物模型是建立人类疾病生物医学模型的基础 以及更好地理解动物生物学。对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的治疗方法至关重要。 使用GE大型动物模型（如猪）通常会产生临床相关结果， 和解剖学上与人类相似例如，引入导致囊性纤维化的遗传因素， 猪基因组的免疫缺陷诱导转基因猪密切重演疾病的症状。然而，在这方面， 由于建立这种模型需要大量时间，转基因猪在生物医学中并不广泛使用。作为 作为一种大型动物物种，猪的单轮育种至少需要一年， 建立转基因猪模型需要育种。基因组编辑工具的应用，如CRISPR/Cas 系统，具有显着提高的功效，引入靶向修饰的猪基因组。然而，在这方面， 对基因组编辑程序引起的非预期基因组改变的担忧以及引入 作为大型动物模型的猪的靶向修饰阻碍了该技术的广泛应用。我们的目标是 该项目旨在评估基因组编辑技术的有效性和安全性，并设计新的方法。 这将有助于在靶向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 是为了实现我们的目标。首先，我们将生成GE中用于全局检测脱靶事件的方法 猪其次，我们将制定策略，以确保基因组编辑过程中的基因组完整性。最后， 我们建议开发一种策略，以快速表型GE胎儿和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的性状相关的基因。 从该项目中获得的知识可以用于扩大转基因猪在生物医学中的应用， 对农业生产也有影响。利用基因组预测表型的重要性 用于快速鉴定改良的等位基因和性状。我们在使用基因组编辑方面的专业知识 技术和GE猪模型将被用来完成所提出的目标。该项目的成果应 通过有效地捕获随后的基因工程猪模型， 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 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， 生物医学和农业上的重要物种。考虑到猪作为动物模型的重要性， 我们的发现对NIH和USDA机构都是有益的。