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Nat Biomed Eng. 2017;1:889-901. doi: 10.1038/s41551-017-0137-2. Epub 2017 Oct 2.

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

Author information

1
GenEdit, Berkeley, CA, 94720-0001, USA.
2
Department of Bioengineering, University of California, Berkeley, Berkeley, CA, 94720, USA.
3
Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA, 94720, USA.
4
Center for Disease Biology and Integrative Medicine, Graduate School of Medicine, The University of Tokyo, Tokyo, 113-0033, Japan.
5
Department of Materials Engineering, Graduate School of Engineering, The University of Tokyo, Tokyo, 113-0033, Japan.
6
Innovative Genomics Initiative, University of California, Berkeley, Berkeley, CA, 94720, USA.
7
Innovation Center of NanoMedicine, Institute of Industry Promotion-KAWASAKI, Kawasaki, 210-0821, Japan.
8
Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, CA, 94720, USA.
9
Department of Chemistry, University of California, Berkeley, Berkeley, CA, 97420-1460, USA.
10
Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA.
11
Department of Bioengineering, University of California, Berkeley, Berkeley, CA, 94720, USA. iconboy@berkeley.edu.
12
Department of Bioengineering, University of California, Berkeley, Berkeley, CA, 94720, USA. nmurthy@berkeley.edu.

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 in vivo 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.

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