Functional Analysis of Genes and Genomic Variants in Animal Models

动物模型中基因和基因组变异的功能分析

基本信息

批准号：
9124899
负责人：
Eric Chien-Wei Liao
金额：
$ 53.9万
依托单位：
BRIGHAM AND WOMEN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2001
资助国家：
美国
起止时间：
2001-04-01 至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/9124899
关键词：
Alleles Anatomy Animal Model Benign Bioinformatics Biological Biological Models Biological Process CRISPR/Cas technology Candidate Disease Gene Clinical Codon Nucleotides Communities Development Disease Expert Systems Functional disorder Gene Structure Gene Targeting Genes Genome Genomics Genotype Goals Human Human Genome Imagery Mediating Methods Modeling Mus Mutagenesis Mutation Orthologous Gene Outcome Pathogenicity Patients Phenotype Point Mutation Reporter Research Research Project Grants Systems Analysis Technology Testing Transgenic Organisms Validation Variant Zebrafish abstracting base biological systems body system clinical phenotype cohort design exome gene discovery gene function genetic variant genome analysis genome editing genome wide association study high throughput analysis high throughput screening human genomics in vitro Model in vivo innovation insight interest knock-down loss of function mutant novel spatiotemporal

项目摘要

PROJECT 2 ABSTRACT/RESEARCH SUMMARY. Project 1 of DGAP takes advantage of a robust clinical referral network and innovations in genome analysis approaches to identify disease associated genes-of-interest using BCAs. Akin to other human genomics research projects (e.g., GWAS, WES/WGS), we are generating a rapidly expanding candidate gene list that is outpacing the functional analysis to validate the causal relationship and understand the underlying pathophysiology. Use of an animal model to disrupt the human ortholog and recapitulate the clinical phenotype remains the best functional evidence for gene function and pathogenicity of gene variants. Major advances in gene targeting methods now make it possible to functionally annotate human candidate genes in a higher throughput and precisely targeted fashion. Further, the biologic advantages and wealth of transgenic reporters allows for rapid phenotypic analysis from direct visualization of the biological process of interest. The goal of Project 2 is to use the convergence of advanced gene editing state-of-the-art biological system analyses and convergent bioinformatics to establish rapidly functional evidence for genes-of-interest. We have designed Project 2 to provide ready-to-use deliverables to the scientific community. Aim 1 utilizes high-throughput in vivo spatiotemporal expression analysis and morpholino-mediated gene knockdown to rapidly analyze the function of candidate genes (approximately 30 per year) discovered at the BCA breakpoints. Aim 2 applies high-throughput targeted mutagenesis using CRISPR/Cas9 approach to generate loss-of-function zebrafish models (approximately 10-15 genes per year) to provide functional evidence for consequences of DGAP gene disruptions in the appropriate biologic context. All organ systems will be examined via rapid phenotype assessment strategy, supported by a consultant team of zebrafish biologists as needed, while genes with potential function in neuro-development will be forwarded for analysis in Project 3. Aim 3 takes advantage of the animal models to carry out high-throughput analysis of human gene variants to determine pathogenicity of the mutation (approximately 10-12 variants per year). Project 2 will make a significant contribution to functional gene annotation discovered through the DGAP pipeline, innovate strategies to analyze human genome variants, and generate deliverables to the scientific community to catalyze biological explorations across organ systems.

项目 2 摘要/研究摘要。 DGAP 项目 1 利用强大的临床转诊网络和基因组分析创新使用 BCA 识别疾病相关感兴趣基因的方法。类似于其他人类基因组学研究项目（例如 GWAS、WES/WGS），我们正在生成一个快速扩展的候选基因列表超越功能分析来验证因果关系并了解根本原因病理生理学。使用动物模型破坏人类直系同源物并重现临床表型仍然是基因功能和基因变异致病性的最佳功能证据。主要进展基因打靶方法现在可以在更高的水平上对人类候选基因进行功能注释吞吐量和精确定位的时尚。此外，转基因报告基因的生物学优势和财富允许通过直接可视化感兴趣的生物过程进行快速表型分析。目标是项目2是利用先进的基因编辑技术与最先进的生物系统的融合分析和融合生物信息学，以快速建立感兴趣基因的功能证据。我们设计项目 2 是为了向科学界提供即用型交付成果。目标1利用高通量体内时空表达分析和吗啉介导的基因敲低快速分析 BCA 发现的候选基因（每年大约 30 个）的功能断点。目标 2 使用 CRISPR/Cas9 方法应用高通量定向诱变来生成功能丧失的斑马鱼模型（每年大约 10-15 个基因）为 DGAP 基因破坏在适当的生物学背景下的后果。所有器官系统都将通过快速表型评估策略进行检查，并得到斑马鱼生物学家顾问团队的支持需要的，而在神经发育中具有潜在功能的基因将被转发到项目 3 中进行分析。目标3利用动物模型对人类基因变异进行高通量分析确定突变的致病性（每年大约 10-12 个变异）。项目2将制作一个对通过 DGAP 管道发现的功能基因注释做出了重大贡献，创新分析人类基因组变异并为科学界提供成果的策略催化跨器官系统的生物探索。