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

基本信息

批准号：
10361239
负责人：
JONATHAN FLINT
金额：
$ 69.93万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-06-18 至 2024-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10361239
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

项目摘要

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 年里，已经鉴定出数千个基因位点，这些基因位点对人类的复杂性状有贡献。啮齿动物，包括常见疾病的模型。研究结果预计将加深我们对疾病和其他具有生物医学意义的特征的生物学机制，但与数量有关成功的绘图实验产生的新颖见解非常少。这是因为映射实验很少导致基因的鉴定。该提案将从根本上改变这种状况采用创新方法来识别基因位点上导致复杂变异的基因小鼠的特征。通过使用 PI 开发的资源和技术，包括使用近交系用于啮齿类动物的高分辨率遗传图谱、基因组中小鼠品系遗传变异的目录测序和基因鉴定方法，将开发一种有效且简单的方案，该方案将使研究人员能够快速从位点鉴定到基因鉴定。由于基因鉴定特别在病因学理解仍然有限的精神疾病模型中很重要（并且具有挑战性）由于难以获取相关组织或细胞类型，因此该方法的功效已在动物模型上进行了测试焦虑。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可靠的；其次，识别每个位点的所有候选基因；第三，在近交系上敲除这些基因并使用数量性状基因座基因敲除相互作用测试来测试他们的候选资格。使用一组发现 62 个基因座导致小鼠焦虑变化，我们的目标是识别 24 个基因座，其中有两个或更少的候选基因基因，并确认这些位点上与焦虑有关的基因的身份。直到最近，关键实验使基因鉴定成为可能的相互作用测试由于很难在相同的遗传背景下获得敲除型和野生型。新事物的出现基因组工程技术 CRISPR/Cas9 克服了这一障碍。我们将利用这一点使复杂性状位点的基因鉴定成为一项常规任务。我们的发现将改变复杂的啮齿类动物的性状遗传学，通过鉴定多达 24 个与焦虑有关的基因，将在了解常见疾病的生物学基础，从而对开发新的疾病产生影响疗法。