Developing a Pathway from Genetic Locus to Gene for Complex Traits in Rodents

开发从遗传位点到啮齿动物复杂性状基因的途径

• 批准号: 10197749
• 负责人: JONATHAN FLINT
• 金额: $ 71.02万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-06-18 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10197749
• 关键词: Alleles; Amygdaloid structure; Analysis of Variance; Animal Model; Anxiety; Biological; CRISPR/Cas technology; Candidate Disease Gene; Catalogs; Chromosome Mapping; Complex; Complex Genetic Trait; Data; Data Set; Development; Disease; Disease model; Engineering; Ensure; Etiology; Functional disorder; Genes; Genetic; Genomics; Genotype; Hippocampus (Brain); Hybrids; Inbred Strain; Inbred Strains Rats; Knock-out; Maps; Medical; Mental disorders; Meta-Analysis; Methods; Modeling; Mouse Strains; Mus; Nucleic Acid Regulatory Sequences; Pathway interactions; Phenotype; Prosencephalon; Protocols documentation; Quantitative Trait Loci; Rattus; Research Personnel; Resolution; Resources; Rodent; Rodent Model; System; Techniques; Technology; Testing; Tissues; Ursidae Family; Variant; causal variant; cell type; experimental study; gene discovery; genetic variant; genome sequencing; genomic locus; ineffective therapies; innovation; insight; interest; knockout gene; model design; mouse genome; mouse model; mutant; novel; novel therapeutics; promoter; success; trait

Over the last 20 years many thousands of genetic loci have been identified that contribute to complex traits in rodents, including models of common diseases. The findings are expected to advance our understanding of biological mechanisms underlying disease and other traits of biomedical interest, yet relative to the number of successful mapping experiments the yield of novel insights is very small. This is because the mapping experiments have rarely led to the identification of genes. This proposal will radically change this situation by deploying an innovative approach to identifying genes at genetic loci that contribute to variation in complex traits in mice. By using resources and techniques that the PI has developed, including the use of outbred rodents for high-resolution genetic mapping, catalogs of genetic variants in mouse strains from genome sequencing, and methods for gene identification, an efficient and simple protocol will be developed that will allow researchers to rapidly progress from locus to gene identification. Since gene identification is particularly important (and challenging) in models of psychiatric disease where etiologic understanding is still limited and access to the relevant tissues or cell type difficult, the efficacy of the approach is tested on animal models of anxiety. Our approach consists of three steps: first, ensure that association evidence supporting each locus is robust; second, identify at each locus all candidate genes; third, make knockouts of those genes on an inbred strain and test their candidacy using a quantitative trait locus gene-knockout interaction test. Using a discovery set of 62 loci that contribute to variation in anxiety in mice, we aim to identify 24 loci with two or fewer candidate genes, and to confirm the identity of genes involved in anxiety at these loci. Until recently the key experiment that makes gene identification possible, the interaction test, could not easily be implemented because of the difficulty of obtaining a knockout and wildtype on the same genetic background. The advent of the new genomic engineering technology, CRISPR/Cas9, has overcome that obstacle. We will take advantage of this advance to make gene identification at complex trait loci a routine task. Our findings will transform complex trait genetics in rodents, and, by identifying up to 24 genes involved in anxiety, will make a major inroad into understanding the biological basis of a common disease, with consequent implications for developing new therapies.

在过去的20年中，已经确定了成千上万的遗传基因座 啮齿动物，包括常见疾病的模型。期望这些发现将提高我们对 生物医学兴趣的生物学机制和其他特征，但相对于数量 成功的映射实验新见解的产量非常小。这是因为映射 实验很少导致基因的鉴定。该建议将通过 部署一种创新的方法来识别遗传基因座的基因，这有助于复杂的变化 小鼠的特征。通过使用PI开发的资源和技术，包括使用杂种 用于高分辨率遗传图的啮齿动物，基因组小鼠菌株中遗传变异的目录 测序和基因识别方法，将开发一种有效而简单的协议，将 允许研究人员快速从基因座到基因鉴定。由于基因鉴定特别是 在精神疾病模型中，病因学仍然有限，并且 难以进入相关的组织或细胞类型，该方法的功效已在动物模型上测试 焦虑。 我们的方法包括三个步骤：首先，确保支持每个基因座的关联证据是强大的； 其次，在每个基因座上识别所有候选基因；第三，将这些基因的敲除近交菌株的敲除 并使用定量性状基因座基因之间的互动测试测试他们的候选人资格。使用一组发现 62个基因座，导致小鼠焦虑症的变化，我们旨在识别24个基因座，以两个或更少的候选者 基因，并确认在这些基因座的焦虑中涉及的基因的身份。直到最近，关键实验 这使得基因识别成为可能，即相互作用测试，因此无法轻易实施 在相同的遗传背景上获得敲除和野生型的困难。新的到来 基因组工程技术CRISPR/CAS9克服了这一障碍。我们将利用这一点 提前使在复杂性状基因座上的基因识别成为常规任务。我们的发现将改变复杂 啮齿动物中的特质遗传学，以及通过识别焦虑涉及的多达24个基因，将大大介入 了解一种常见疾病的生物学基础，因此对发展新的疾病产生了影响 疗法。