CRISPR-Cas9 Gene Editing Protects from the A53T-SNCA Overexpression-Induced Pathology of Parkinson's Disease <i>In Vivo</i>

Abstract

Mutations in specific genes, including synuclein alpha (<i>SNCA</i>) that encodes the α-synuclein protein, are known to be risk factors for sporadic Parkinson's disease (PD), as well as critical factors for familial PD. In particular, A53T-mutated <i>SNCA</i> (A53T-SNCA) is a well-studied familial pathologic mutation in PD. However, techniques for deletion of the mutated <i>SNCA</i> gene <i>in vivo</i> have not been developed. Here, we used the CRISPR-Cas9 system to delete A53T-SNCA <i>in vitro</i> as well as <i>in vivo</i>. Adeno-associated virus carrying SaCas9-KKH with a single-guide RNA targeting A53T-SNCA significantly reduced A53T-SNCA expression levels <i>in vitro</i>. Furthermore, we tested its therapeutic potential <i>in vivo</i> in a viral A53T-SNCA-overexpressing rat model of PD. Gene deletion of A53T-SNCA significantly rescued the overexpression of α-synuclein, reactive microgliosis, dopaminergic neurodegeneration, and parkinsonian motor symptoms. Our findings propose CRISPR-Cas9 system as a potential prevention strategy for A53T-SNCA-specific PD.