Efficient Gene Editing through Direct Cytosolic Delivery of CRISPR/Cas9-Ribonucleoprotein

Genome editing through the delivery of CRISPR/Cas9-ribonulceoprotein (Cas9-RNP) reduces unwanted gene targeting and avoids integrational mutagenesis that can occur through gene delivery strategies. Direct and efficient delivery of Cas9-RNP into the cytosol followed by translocation to the nucleus remains a challenge. Here we report a remarkably high efficient (~90%) direct cytoplasmic/nuclear delivery of Cas9 protein complexed with a guide RNA (sgRNA) through the co-engineering of Cas9 protein and carrier nanoparticles. This construct provides effective (~30%) gene editing efficiency and opens up new opportunities in studying genome dynamics.