Self-Assembled DNA Nanoclews for the Efficient Delivery of CRISPR-Cas9 for Genome Editing

CRISPR-Cas9 represents a promising technology for genome editing, yet means of safe and efficient delivery remain to be fully realized. Here, we report a novel delivery vehicle to deliver the Cas9 protein and single-guide RNA simultaneously based on DNA nanoclews, yarn-like DNA nanoparticles synthesized by rolling circle amplification. The bio-inspired vehicles efficiently loaded Cas9/single-guide RNA complexes and delivered the complexes to the nuclei of human cells, allowing targeted gene disruptions while maintaining cell viability. Editing was most efficient when the DNA nanoclew sequence and the sgRNA guide sequence were partially complementary, offering a design rule for enhancing delivery. Overall, this strategy provides a versatile platform that could be adapted for delivering other DNA-binding proteins or for functional nucleic acids