CRISPR/Cas9-mediated correction of mutations that cause inherited retinal degenerations

CRISPR/Cas9介导的导致遗传性视网膜变性的突变的校正

基本信息

批准号：
9238770
负责人：
EDWIN M STONE
金额：
$ 34.31万
依托单位：
UNIVERSITY OF IOWA
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-04-01 至 2019-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9238770
关键词：
Adaptive Immune System Address Affect Alleles Animal Model Autologous Transplantation Cells Cessation of life Code Codon Nucleotides Coupling Development Disease Future GTP-Binding Protein alpha Subunits, Gs Gene Targeting Generations Generic Drugs Genes Genetic Genomics Goals Guide RNA Human In Vitro Individual Inherited Light Mediating Miniature Swine Modality Molecular Mutation Neural Retina Nonhomologous DNA End Joining Pathogenicity Patients Phenotype Photoreceptors Prevalence RNA Splicing Reagent Reporting Retina Retinal Dystrophy Retinitis Pigmentosa Site Splice-Site Mutation Stem cells System Testing Therapeutic Toxic effect Transgenic Organisms Usher Syndrome, Type 2A Variant Viral base clinical phenotype disease phenotype disease-causing mutation gain of function gain of function mutation gene repair gene replacement therapy genome editing in vivo induced pluripotent stem cell inherited retinal degeneration loss of function mutation nuclease overexpression precursor cell promoter public health relevance repaired treatment strategy

项目摘要

DESCRIPTION (provided by applicant): Retinitis pigmentosa (RP) is a genetically heterogeneous disorder characterized by death of the light-sensing photoreceptor cells of the outer neural retina. Although collectively quite common, individually each disease- causing gene is quite rare. Three genes, USH2A, ABCA4 and RHO, account for a large portion of disease and elicit a wide-range of RP clinical phenotypes caused by an array of different disease-causing mutations. In this proposal we will use CRISPR/Cas9-mediated genome editing to target and correct three different classes of disease-causing mutations: 1) an intronic cryptic splice site mutation in USH2A; 2) an exonic coding sequence mutation in ABCA4; and 3) a dominant gain-of-function mutation in RHO. Using a CRISPR-based strategy will allow for disease-causing genes to remain under control of their endogenous promoters, allow for correction of large genes not amenable to gene replacement therapy and provide an avenue to `turn off' harmful dominant gain of function mutations. These genome-editing approaches will be tested on patient-derived induced pluripotent stem cells with molecularly confirmed mutations. Correcting cells that harbor a disease-causing mutation will allow for generation of cells for future autologous transplantation studies in human patients. The studies outlined in this proposal will pave the way for a new gene augmentation and stem cell-based therapeutic avenues for treatment of retinitis pigmentosa.

描述（由适用提供）：色素性视网膜炎（RP）是一种遗传异质性疾病，其特征是外神经视网膜的光感应感光细胞死亡。尽管总体上很普遍，但单独的每个疾病 - 引起基因很少见。三个基因USH2A，ABCA4和RHO占了很大一部分疾病，并引起了由一系列由不同的引起疾病的突变引起的大量RP临床表型。在此提案中，我们将使用CRISPR/CAS9介导的基因组编辑来靶向并纠正三种不同类别的致病突变：1）USH2A中的内含子加密剪接位点突变； 3）RHO中的主要功能收益突变。使用基于CRISPR的策略将使引起疾病的基因保持其内源性启动子的控制，允许校正不适合基因替代疗法的大基因，并提供了“关闭”有害的功能突变获得的途径。这些基因组编辑的方法将对具有分子确认的突变的患者衍生诱导的多能干细胞进行测试。纠正携带引起疾病突变的细胞将允许在人类患者中生成未来自体移植研究的细胞。该提案中概述的研究将为新的基因增强和基于干细胞的治疗途径铺平道路。