Nanoparticle delivery of Cas9 ribonucleoprotein and donor DNA <i>in vivo</i> induces homology-directed DNA repair.

Lee, Kunwoo; Conboy, Michael; Park, Hyo Min; Jiang, Fuguo; Kim, Hyun Jin; Dewitt, Mark A; Mackley, Vanessa A; Chang, Kevin et al. · Nat Biomed Eng · 2017

basic_science · Level V

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Abstract

CRISPR/Cas9-based therapeutics, especially those that can correct gene mutations via homology directed repair (HDR), have the potential to revolutionize the treatment of genetic diseases. However, HDR-based therapeutics are challenging to develop because they require simultaneous <i>in vivo</i> delivery of Cas9 protein, guide RNA and donor DNA. Here, we demonstrate that a delivery vehicle composed of gold nanoparticles conjugated to DNA and complexed with cationic endosomal disruptive polymers can deliver Cas9 ribonucleoprotein and donor DNA into a wide variety of cell types, and efficiently correct the DNA mutation that causes Duchenne muscular dystrophy in mice via local injection, with minimal off-target DNA damage.