A general genome editing strategy using CRISPR lipid nanoparticle spherical nucleic acids

Sep 4, 2025Proceedings of the National Academy of Sciences of the United States of America

A general method for genome editing using CRISPR delivered by lipid nanoparticle spheres

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Abstract

CRISPR lipid nanoparticle-spherical nucleic acids (LNP-SNAs) demonstrate a 2.5-fold improvement in homology-directed repair efficiency compared to standard lipid nanoparticles.

CRISPR LNP-SNAs achieve two- to three-fold higher cellular uptake than lipid nanoparticles without a DNA shell.
Reduced cytotoxicity is observed with CRISPR LNP-SNAs compared to conventional delivery vehicles.
Gene transfection efficiency with CRISPR LNP-SNAs is also two- to three-fold greater than that with standard lipid nanoparticles.
Insertion-deletion mutations are induced at average frequencies two- to three-fold higher with CRISPR LNP-SNAs than with traditional lipid nanoparticles.
The combination of two nucleic acid delivery approaches enhances the potential of CRISPR LNP-SNAs for gene therapy applications.

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Genome editing with CRISPR-Cas systems hold promise for treating a wide range of genetic disorders and cancers. However, efficient delivery of genome editors remains challenging due to the requirement for the simultaneous delivery or intracellular generation of Cas proteins, guide RNAs, and, in some applications, donor DNAs. Furthermore, the immunogenicity and toxicity of delivery vehicles can limit the safety and efficacy of genetic medicines. Here, we combine two nucleic acid delivery approaches to create CRISPR lipid nanoparticle-spherical nucleic acids (LNP-SNAs) that are both efficient and biocompatible. Compared to lipid nanoparticles (LNPs) lacking a surface-bound DNA shell, CRISPR LNP-SNAs exhibit two- to three-fold higher cellular uptake, reduced cytotoxicity, and improved gene transfection efficiency. Across multiple cell lines and genomic loci, CRISPR LNP-SNAs induce insertion-deletion mutations at average frequencies two- to three-fold higher than those observed with LNPs. When codelivered with donor templates, CRISPR LNP-SNAs enable homology-directed repair at an average efficiency of 21 ± 7%, a 2.5-fold improvement over LNPs (8 ± 4%). The ease of synthesis and biocompatibility of CRISPR LNP-SNAs highlight their potential as a versatile delivery platform for CRISPR-Cas and other gene therapies.