Develop combinatorial non-viral and viral CRISPR delivery for lung diseases

开发针对肺部疾病的组合非病毒和病毒 CRISPR 递送

基本信息

批准号：
10274832
负责人：
DANIEL G ANDERSON
金额：
$ 127.79万
依托单位：
UNIV OF MASSACHUSETTS MED SCH WORCESTER
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：
2018-09-10 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10274832
关键词：
Adult Animal Model Bar Codes Basic Science Biotechnology CRISPR therapeutics CRISPR/Cas technology Capsid Cells Clustered Regularly Interspaced Short Palindromic Repeats Cystic Fibrosis Cystic Fibrosis Transmembrane Conductance Regulator DNA DNA Sequence Delta F508 mutation Dependovirus Development Disease Dose Epithelial Cells Evaluation Future Gene Delivery Gene Mutation Generations Genes Genetic Genetic Diseases Goals Guide RNA Length Libraries Liver Lung Lung diseases Macaca Macaca mulatta Measures Mediating Messenger RNA Modeling Mus Mutation Nature Nebulizer Nonhomologous DNA End Joining Nonsense Mutation Pan Genus Pharmaceutical Preparations Phase Polymers Primates Production Reporter Rhesus Risk Safety Serotyping Specificity Structure System Technology Testing Therapeutic Human Experimentation Therapeutic Studies Toxic effect Variant Viral Viral Genome Work Yin airway epithelium arm cell type combinatorial deep sequencing design gene correction gene therapy genome editing high throughput screening homologous recombination improved in vivo in vivo Model innovation insertion/deletion mutation lung repair nanoparticle nanoparticle delivery nonhuman primate novel nuclease protein expression repaired scale up somatic cell gene editing stem cells targeted delivery therapeutic genome editing therapeutic protein tool

项目摘要

Project Summary The ability to correct disease gene mutations in vivo has broad potential utility for both therapy and basic research. CRISPR/Cas9 is a powerful RNA-guided tool for genome editing. Our recent discovery that CRISPR/Cas9 delivery can cure genetic disease in adult mouse liver provided proof-of-concept of gene correction therapy in vivo. The main goal of this proposal is to establish innovative delivery technologies to maximize the efficiency of CRISPR delivery and gene correction in disease-relevant lung cell types. The impact of this project is a novel paradigm of lung-targeted delivery tools for CRISPR-mediated gene correction. The development of safe and effective delivery vehicles and genome editing tools will guide future studies for CRISPR-mediated gene therapy. This project has three aims that focus on different aspects of lung-directed somatic genome editing: Aim 1: Optimize NP+AAV combination for delivery and gene correction in mouse lung; Aim 2: Identify the best AAV capsid for lung delivery and optimize AAV genome as donor template for HDR; Aim 3: Characterize NP+AAV developed in the UG3 phase in macaque models. This project will develop innovative designs of adeno-associated virus and nanoparticles to significantly improve the efficiency, cell specificity, and safety of CRISPR delivery in vivo.

项目摘要在体内纠正疾病基因突变的能力具有两种治疗的广泛潜在效用和基础研究。 CRISPR/CAS9是用于基因组编辑的强大RNA引导的工具。我们的最近发现CRISPR/CAS9的递送可以治愈成年小鼠肝的遗传疾病提供了体内基因矫正疗法的概念证明。该提议的主要目标是建立创新的交付技术，以最大化CRISPR交付的效率和与疾病相关的肺部细胞类型的基因校正。这个项目的影响是一本小说针对CRISPR介导的基因校正的肺靶向输送工具的范式。这开发安全有效的送货工具以及基因组编辑工具将指导未来 CRISPR介导的基因疗法的研究。该项目的三个目标专注于不同肺导向的体细胞基因组编辑的各个方面：目标1：优化NP+AAV组合小鼠肺的递送和基因校正； AIM 2：确定最佳的AAV CAPSID 输送并优化AAV基因组作为HDR的供体模板； AIM 3：表征NP+AAV 在猕猴模型中在UG3阶段开发。该项目将开发出创新的设计腺相关病毒和纳米颗粒可显着提高效率，细胞特异性，和CRISPR在体内交付的安全。