A CRISPR-engineered swine model of COL2A1 deficiency recapitulates altered early skeletal developmental defects in humans.

Loss-of-function mutations in the COL2A1 gene were previously described as a cause of type II collagenopathy (e.g., spondyloepiphyseal dysplasia, Stickler syndrome type I), a major subgroup of genetic skeletal diseases. However, the pathogenic mechanisms associated with COL2A1 mutations remain unclear, and there are few large-mammal models of these diseases. In this study, we established a swine model carrying COL2A1 mutations using CRISPR/Cas9 and somatic cell nuclear transfer technologies. Animals mutant for COL2A1 exhibited severe skeletal dysplasia characterized by shortened long bones, abnormal vertebrae, depressed nasal bridge, and cleft palate. Importantly, COL2A1 mutant piglets suffered tracheal collapse, which was almost certainly the cause of their death shortly after birth. In conclusion, we have demonstrated for the first time that overt and striking skeletal dysplasia occurring in human patients can be recapitulated in large transgenic mammals. This model underscores the importance of employing large animals as models to investigate the pathogenesis and potential therapeutics of skeletal diseases.