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

TLDR

CRISPR–Cas9 is a promising genome editing platform, but safe and efficient delivery methods are still lacking. The authors developed DNA nanoclews—yarn‑like nanoparticles produced by rolling circle amplification—that load Cas9/sgRNA complexes and deliver them to human cell nuclei for targeted gene disruption without compromising viability. Editing efficiency was highest when the nanoclew and sgRNA sequences were partially complementary, providing a design rule, and the approach is versatile for delivering other DNA‑binding proteins or nucleic acids.

Abstract

Abstract CRISPR–Cas9 represents a promising platform for genome editing, yet means for its safe and efficient delivery remain to be fully realized. A novel vehicle that simultaneously delivers the Cas9 protein and single guide RNA (sgRNA) is based on DNA nanoclews, yarn‐like DNA nanoparticles that are synthesized by rolling circle amplification. The biologically inspired vehicles were efficiently loaded with Cas9/sgRNA complexes and delivered the complexes to the nuclei of human cells, thus enabling targeted gene disruption 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 method that could be adapted for delivering other DNA‐binding proteins or functional nucleic acids.