Reduced Off-Target Effects with Delivery of Active Cas9 Protein Complexed with sgRNA Using Gesicle Technology

The Guide-it CRISPR/Cas9 Gesicle Production System is a novel methodology that uses cell-derived nanovesicles, called gesicles, for co-delivery of active Cas9 protein complexed with a gene-specific sgRNA. Gesicles are loaded with Cas9-sgRNA ribonucleoprotein (RNP) complexes, and can be added directly to target cells for CRISPR/Cas9 gene editing. Delivery of active Cas9 protein means no Cas9 coding gene is present in target cells, thus eliminating the problem of persistent and elevated of Cas9, which is common to plasmid-based delivery methods. This technical note highlights a side-by-side comparison of Cas9-sgRNA delivery methods.