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.