Systemic CRISPR Therapy for the Treatment of Inherited Diseases

This technology includes novel systemic adeno-associated virus (AAV)-mediated CRISPR gene therapy technology. While some diseases (e.g., retinal diseases) can be treated through local gene transfer, many diseases such as Duchenne Muscular Dystrophy (DMD) require systemic therapy. The CRISPR technology has two components, the Cas9 endonuclease, and the gRNA. To explore systemic CRISPR therapy, we co-delivered the AAV.Cas9 and AAV.gRNA vector to mdx mice, a mouse DMD model. Direct delivery to muscle yielded efficient gene correction. Through detailed analysis of gene editing at the protein and transcript level and detailed profiling of the two AAV CRISPR vectors, we identified preferential loss of the gRNA vector as the barrier to systemic AAV CRISPR therapy. Increasing gRNA vector dose successfully overcome this hurdle and resulted in correction in both heart and skeletal muscle. Importantly, modified CRISPR therapy significantly enhanced skeletal muscle and cardiac function. The patent illustrates the development of this novel systemic AAV CRISPR gene therapy technology, which can be applied to treat many inherited diseases.