Non-viral Delivery of CRISPR/Cas9 Gene Editing for Angelman Syndrome

With a CRISPR/Cas9 genome editing tool, this project aims to restore UBE3A expression from the paternal chromosome via a single treatment approach. The paternal UBE3A allele is silenced by a long noncoding RNA that is antisense to UBE3A. This strategy will unsilence the paternal UBE3A allele, using CRISPR/Cas9 genome editing. This treatment will be delivered through an innovative nonviral chemically modified ribonucleoprotein (RNP) complex (cRNP-Cas9/gRNA) delivery system. It will carry the ready Cas9 protein and single guide RNA in one intrathecal injection to permanently activate the paternal UBE3A expression. The proposed study will not only provide landmark knowledge on non-viral delivery systems of CRISPR/Cas9 gene therapy, but also support the possibility of translational application using this system as a therapeutic approach to treat patients with Angelman syndrome.