Subject(s)
Cartilage, Articular/drug effects , Chondrocytes/drug effects , Glucuronidase/therapeutic use , Osteoarthritis, Knee/drug therapy , Receptor, Transforming Growth Factor-beta Type II/therapeutic use , Animals , Cartilage, Articular/pathology , Cell Culture Techniques , Chondrocytes/pathology , Humans , Klotho Proteins , Osteoarthritis, Knee/chemically induced , Papain , Rats , Recombinant Proteins/therapeutic useABSTRACT
In vivo genome editing represents a powerful strategy for both understanding basic biology and treating inherited diseases. However, it remains a challenge to develop universal and efficient in vivo genome-editing tools for tissues that comprise diverse cell types in either a dividing or non-dividing state. Here, we describe a versatile in vivo gene knock-in methodology that enables the targeting of a broad range of mutations and cell types through the insertion of a minigene at an intron of the target gene locus using an intracellularly linearized single homology arm donor. As a proof-of-concept, we focused on a mouse model of premature-aging caused by a dominant point mutation, which is difficult to repair using existing in vivo genome-editing tools. Systemic treatment using our new method ameliorated aging-associated phenotypes and extended animal lifespan, thus highlighting the potential of this methodology for a broad range of in vivo genome-editing applications.