Fluoropolymer‐Mediated Delivery of a Dual TSHR/IGF1R‐Targeting CRISPR‐Cas9 System for Localized Therapy in Thyroid‐Associated Ophthalmopathy

ABSTRACT Thyroid‐associated ophthalmopathy (TAO), a vision‐threatening and disfiguring autoimmune orbital disorder, remains a therapeutic challenge due to the lack of therapies with orbital specificity, sustained efficacy, and minimal side effects. Herein, we present G4F7‐CRISPR, a fluoropolymer‐based CRISPR‐Cas9 delivery platform engineered for localized and efficient disruption of thyroid‐stimulating hormone receptor ( TSHR ) and insulin‐like growth factor 1 receptor ( IGF1R ), two key mediators of TAO pathogenesis. G4F7‐CRISPR achieved high insertion/deletion frequencies in primary orbital fibroblasts ( Tshr : 37.2%; Igf1r : 42.8%) and mature adipocytes ( Tshr : 22.4%; Igf1r : 24.3%), and maintained robust editing efficiency in orbital adipose tissue of TAO mouse models ( Tshr : 30.7%; Igf1r : 32.4%). In both TAO mouse models and 3D human orbital organoids, dual‐gene editing of Tshr and Igf1r via G4F7‐CRISPR significantly suppressed orbital adipogenesis, inflammation, and fibrosis, demonstrating superior therapeutic efficacy over either single‐gene approaches. Comprehensive off‐target analyses in both TAO mouse models and orbital organoids revealed minimal off‐target activity. Furthermore, G4F7‐CRISPR exhibited excellent short‐ and long‐term ocular and systemic safety in TAO mouse models. Notably, it outperformed teprotumumab—the FDA‐approved therapy for TAO—in both therapeutic efficacy and safety, highlighting its potential clinical advantages. Collectively, these findings highlight the translational promise of G4F7‐CRISPR as a safe, precise, and clinically viable gene therapy for TAO.