CRISPR/Cas9-mediated gene correction in newborn rabbits with hereditary tyrosinemia type I

Abstract Patients with hereditary tyrosinemia type I (HT1) present acute and irreversible liver and kidney damages during infancy. CRISPR/Cas9-mediated gene correction during infancy may provide a promising approach to treat patients with HT1. However, all previous studies were performed on adult HT1 rodent models, which cannot authentically recapitulate some symptoms of human patients. To translate precise gene therapy to clinical practice, the efficacy and safety should be verified in large animals. Here, we delivered CRISPR/Cas9 and donor templates via adeno-associated virus to newborn HT1 rabbits. The lethal phenotypes could be rescued, and notably, these HT1 rabbits reached adulthood normally without 2-(2-nitro-4-trifluoromethylbenzyol)-1,3 cyclohexanedione administration, and even gave birth to offspring. AAV-treated HT1 rabbits displayed normal liver and kidney structures and functions. Homology-directed repair-mediated precise gene corrections and non-homologous end joining-mediated out-of-frame to in-frame corrections in the livers were observed with efficiencies of 0.90%–3.71% and 2.39%–6.35%, respectively, which appeared to be sufficient to recover liver function and decrease liver and kidney damages. This study provides useful large-animal preclinical data for rescuing hepatocyte-related monogenetic metabolic disorders with precise gene therapy.