Liver-directed somatic gene correction rAAV system of regulatable Cas9/sgRNA

可调节Cas9/sgRNA的肝脏定向体细胞基因校正rAAV系统

基本信息

批准号：
9071194
负责人：
Wen Xue
金额：
$ 50.25万
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：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/9071194
关键词：
Adult Alleles Animal Model Basic Science Biotechnology CRISPR/Cas technology Capsid Clinical Clinical Trials Clustered Regularly Interspaced Short Palindromic Repeats Development Disease Future Gene Mutation Genes Glutamates Goals Guide RNA Hepatocyte Hereditary Disease Homologous Gene Human Knock-in Mouse Laboratories Length Leukocyte Elastase Liver Lung Lung diseases Lysine Maps Mediating Methods Molecular Mus Mutation Nature Nonhomologous DNA End Joining Patients Phenotype Point Mutation Production Protease Inhibitor Pulmonary Emphysema RNA Interference Recombinant adeno-associated virus (rAAV)Respiratory physiology Safety Serotyping Serum Somatic Cell Staphylococcus aureus System Testing Therapeutic Therapeutic Studies Tissues Translating Work Yin abstracting adeno-associated viral vector alpha 1-Antitrypsin alpha 1-Antitrypsin Deficiency arm clinical application clinically relevant deep sequencing design gene correction gene function gene therapy genome editing homologous recombination improved in vivo innovation knock-down mouse model mutant novel novel strategies pre-clinical preclinical study repaired restoration tool vector

项目摘要

Project Summary/Abstract – Project 3: 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. This subproject will interact synergistically with all other Projects and Cores of this tPPG to develop new rAAV CRISPR tools to treat alpha- 1 antitrypsin deficiency (AATD). The main goal of this proposal is to establish a pre-clinical rAAV paradigm for CRISPR-mediated correction of AAT deficiency in mouse models carrying the mutant human AAT gene. The impact of this project is to develop somatic gene correction using rAAV systems to (1) maximize efficiency and safety of CRISPR delivery, (2) maximize the rate of homologous recombination for gene correction, and (3) efficiently correct AAT mutation in the liver to treat lung disease in mice. The development of safe and effective delivery vehicles and genome editing tools to correct AAT deficiency will guide future clinical trials for CRISPR- mediated gene therapy for AAT lung disease. Because AAV serotypes can target a wide range of tissues, our approach has broad basic research and clinical applications beyond AATD. Project 3 has three Aims that focus on different aspects of liver-directed somatic AAT correction: Aim 1: Develop liver-directed rAAV vehicles to maximize efficiency and precision of Z-AAT genome-editing in mice. Aim 2: Investigate rAAV HDR templates to correct Z-AAT mutation in mouse liver. Aim 3: Explore Z-AAT correction in mouse models in vivo to treat the lung disease.

项目摘要/摘要 – 项目 3：体内纠正疾病基因突变的能力对于两者都具有广泛的潜在用途 CRISPR/Cas9 是一种强大的 RNA 引导基因组编辑工具。我们最近发现 CRISPR/Cas9 递送可以治愈成年小鼠肝脏的遗传性疾病提供了基因校正疗法的概念验证。该子项目将协同相互作用。与该 tPPG 的所有其他项目和核心一起开发新的 rAAV CRISPR 工具来治疗 α- 1 抗胰蛋白酶缺乏症（AATD）该提案的主要目标是建立临床前 rAAV。 CRISPR介导的纠正携带AAT缺陷的小鼠模型的范例该项目的影响是利用突变的人类AAT基因来开发体细胞基因校正。 rAAV 系统 (1) 最大限度地提高 CRISPR 递送的效率和安全性，(2) 最大限度地提高速率用于基因校正的同源重组，以及（3）有效校正AAT突变开发安全有效的递送载体和治疗小鼠肺部疾病。纠正 AAT 缺陷的基因组编辑工具将指导未来的 CRISPR 临床试验因为 AAV 血清型可以针对广泛的范围。除了 AATD 之外，我们的方法还具有广泛的基础研究和临床应用。项目 3 有三个目标，重点关注肝脏定向体细胞 AAT 校正的不同方面：目标 1：开发肝脏导向的 rAAV 载体，最大限度地提高 Z-AAT 的效率和精度目标 2：研究 rAAV HDR 模板以纠正小鼠中的 Z-AAT 突变。目标 3：探索小鼠模型中的 Z-AAT 校正以治疗肺部疾病。