Abstract

Targeting Viral Genomic RNA by the Delivery of Extracellular Vesicles-Mediated CRISPR Machinery

Houjian Cai, Associate Professor, University of Georgia

Extracellular vesicles (EVs) have recently been co-opted as vehicles for the delivery of therapeutics, including CRISPR-Cas9 (Cas9), and are now being modified for higher gene editing efficiency. N-myristoylation is known to translocate Src kinase to the membrane. We reasoned that fusion of the N-terminal of Src to Cas9 may increase localization with the membrane, and subsequently increase EV-loading and gene editing efficacy in EV-treated recipient cells. Our study demonstrate that fusion of the octapeptide to Cas9 induced N-myristoylation and encapsulation of the mCas9/sgRNA complex into EVs. We provide proof of concept for N-myristoylation as a method to increase EV-mediated delivery of therapeutics. The technology can be applied for targeting oncogenic genes in prostate cancer cells.