Development of tools for site-directed analysis of gene function

基因功能定点分析工具的开发

基本信息

批准号：
10187374
负责人：
KARL J CLARK
金额：
$ 6.23万
依托单位：
IOWA STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-15 至 2021-08-14
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10187374
关键词：
AGFG1 gene Actins Address Adhesions Adopted Adult Affect Alleles Animal Model Animals Annexins Binding Biological Blood Vessels Blood flow CRISPR/Cas technology Carrier Proteins Catalogs Cell Communication Cell Proliferation Cell membrane Cells Complex Custom Cytoskeleton DNA Double Strand Break Data Defect Development Disease Disease Progression Endothelial Cells Endothelium Event Female Fetal Development Financial compensation Gene Delivery Gene Expression Gene Family Gene Mutation Genes Genetic Genetic Polymorphism Genetic Transcription Genome Goals Growth Health Health Promotion Hour Human Hypoxia KDR gene Knock-in Knock-out Lesion Malignant Female Reproductive System Neoplasm Malignant Neoplasms Malignant neoplasm of ovary Maternal Mortality Mediating Methodology Methods Modeling Molecular Morphology Mutation Nucleotides Operating System Organ Outcome Ovary Pathologic Neovascularization Pathology Patients Periodicity Phenocopy Phenotype Physiologic Neovascularization Placenta Polycystic Ovary Syndrome Pregnancy Process Publishing RNA Splicing Reproductive Health Role Scaffolding Protein Signal Transduction Signaling Molecule Site Site-Directed Mutagenesis System Techniques Technology Testing Texas Time Tissues Transcript Uterine hemorrhage Variant Woman Writing Zebrafish angiogenesis beta catenin cadherin 5 cancer diagnosis cell motility corpus luteum design endometriosis female reproductive system gene function human disease human model in vivo insertion/deletion mutation interest knock-down malignant breast neoplasm membrane activity mutant new growth nuclease reproductive function reproductive organ response tool tool development transcription activator-like effector nucleases wound healing

项目摘要

The overarching goal of this application is to create tools and efficient methods to define genes that can promote human health. While a tremendous amount of data has been cataloged on gene mutation and changes in gene expression associated with complex human disease, our understanding of those genes that could be co-opted to restore patient health is lacking. To address this need and test for genes that when restored to wild type function promote health, we propose develop mutagenic, revertible and conditional alleles that provide spatial and temporal control of gene expression. The ability to make site-specific, untagged mutant alleles in zebrafish and other models has been greatly advanced by custom nucleases that include TALENs and CRISPR/Cas9 systems. These systems operate on the same principle: they are designed to bind to specific sequences in the genome and create a double strand break. The goals of this proposal leverage the activities of TALEN and CRISPR/Cas9 technologies to make site-specific double strand breaks. These tools and techniques will have direct implications for providing precise gene editing techniques to assess the roles of genes in disease and their ability to promote health following disease progression. While we will develop these methodologies in zebrafish due to their ease of gene delivery, we anticipate these methodologies will not only enhance the efficiency of gene editing but will be readily adaptable for use in other model organisms and large animals. In our opinion, this will have important implications for modeling human disease and health in animal systems by greatly enhancing the ability to make predictable alleles, small nucleotide polymorphisms similar to those associated with human disease, and conditional alleles to test for the ability of a gene to restore health.

此应用程序的总体目标是创建工具和有效的方法来定义可以促进人类健康的基因。虽然已将大量数据分类关于基因突变以及与复杂人类疾病相关的基因表达的变化，我们对那些可以选择恢复患者健康的基因的理解缺乏。到满足这种需求并测试基因，这些基因恢复到野生型功能会促进健康，我们提出发展诱变，可逆和有条件的等位基因，以提供空间和基因表达的时间控制。制造特定地点，未标记的突变等位基因的能力在斑马鱼和其他模型中，通过包括 Talens和CRISPR/CAS9系统。这些系统以相同的原则运行：它们是设计用于与基因组中的特定序列结合并产生双链断裂。这该建议的目标利用Talen和Crispr/Cas9技术的活动来制作特定于现场的双链断裂。这些工具和技术将对提供精确的基因编辑技术来评估基因在疾病及其中的作用疾病进展后促进健康的能力。虽然我们会开发这些斑马鱼的方法学的方法由于其易于基因递送，我们预料到这些方法论不仅会提高基因编辑的效率，而且很容易适应用于其他模型生物和大型动物。我们认为，这将具有重要的通过大大增强动物系统中的人类疾病和健康的影响具有可预测等位基因的能力，小核苷酸多态性类似于相关的核苷酸有人类疾病和有条件的等位基因测试基因恢复健康的能力。