A Community Zebrafish Resource for Modeling GWAS Biology

项目总结： 我们的建议“社区斑马鱼资源用于模拟GWAS生物学”应用了新的方法， 在我们联盟内开发的算法和方法，用于使用以下技术对人类GWASHITS进行功能分析 斑马鱼模型系统。我们的联合体在斑马鱼遗传学、基因组学方面积累了丰富的经验 编辑、生物信息学、功能分析开发、人类疾病建模以及创新 阐明个体基因功能的机械论研究。在前一个周期的奖项中，我们 斑马鱼编码序列变异的高效高含量功能分析平台的建立 以补充Gwas项目，涵盖各种复杂的人类特征，并提供途径切入点 以及了解疾病的遗传模型。在这个续订应用中，我们扩展了我们的功能探索 单个和/或多个GWAS基因座及其在人类医学特征潜在的生物网络中的作用 通过以下具体目标包括普遍存在的非编码变异和药物反应： 目标1-从功能上分析血液、肝脏、心脏和血管特征的多个GWAS研究中的基因座，优化 利用CRISPR-Cas9技术对斑马鱼进行检测和基因编辑。 A)使用经过验证的分析方法，我们将检查GWAS基因座中多个编码基因的功能 使用CRISPR-Cas9技术在斑马鱼中建立遗传模型。 B)通过优化斑马鱼基因组编辑，加速体内基因功能评估 CRISPR-CAS9技术。 目的2-调查斑马鱼不同细胞系中基因组调节区的情况，并使用 信息，以支持更好的地图绘制和功能评估的调节变异来自GWA基因座。 A)利用ATAC-SEQ、CHIP-SEQ、甲基-SEQ和伴随的RNA-SEQ分析，我们将建立一个 不同细胞系和器官中保守(同源)调节区的综合数据库 斑马鱼和人类都是。我们将在不同发育阶段的不同细胞类型中定义调节区 斑马鱼细胞谱系分化、维持和功能的重要阶段。 B)我们将把Gwas基因座的调控效应映射到受保护的斑马鱼调控区域，并测试 利用基因组编辑对其在特定GWA型性状中的作用产生的个体和/或多重调控效应 斑马鱼的技术和经过验证的功能分析。 C)根据功能和编目基因组编辑试剂对保守的Gwas基因座进行分类 疾病建模、机理研究和药物发现。

PROJECT SUMMARY: Our proposal “Community Zebrafish Resource for Modeling GWAS Biology” applies novel approaches, algorithms and methods developed within our consortium for functional analysis of human GWAS hits using the zebrafish model system. Our consortium has accumulated broad experience in zebrafish genetics, genome editing, bioinformatics, functional assay development, and human disease modeling as well as innovative mechanistic studies to elucidate individual gene function. During the previous cycle of the award, we established efficient high-content platforms for the functional analysis of coding sequence variation in zebrafish to complement GWAS projects across a variety of complex human traits, and provided pathway entry points and genetic models for understanding disease. In this renewal application, we extend our functional exploration of single and/or multiple GWAS loci and their roles in biological networks underlying human medical traits to include prevalent non-coding variation and drug responses through the following Specific Aims: Aim 1-Functionally analyze loci from multiple GWAS studies on blood, liver, heart and vessel traits, optimizing assay development and gene editing using CRISPR-Cas9 technology in zebrafish. a) Using validated assays we will examine the function of multiple coding genes in GWAS loci and generate genetic models in the zebrafish using CRISPR-Cas9 technology. b) Accelerate in vivo gene function evaluation by optimization of genome editing in zebrafish using the CRISPR-Cas9 technology. Aim 2-Survey landscapes of genomic regulatory regions in different cell lineages in zebrafish and use the information to support better mapping and functional evaluation of regulatory variants from GWAS loci. a) Using ATAC-seq, ChIP-seq, Methyl-seq, and companion RNA-seq analyses we will establish a comprehensive database of conserved (orthologous) regulatory regions in different cell lineages and organs of both zebrafish and human. We will define regulatory regions in different cell types at different developmental stages which are important for cell lineage differentiation, maintenance and function in zebrafish. b) We will map regulatory effects from GWAS loci to conserved zebrafish regulatory regions and test individual and/or multiple regulatory effects for their roles in specific GWAS traits using genome editing technology and validated functional assays in zebrafish. c) Categorize conserved GWAS loci based on function and catalog genome editing reagents for disease modeling and mechanistic studies and drug discovery.