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Nat Biotechnol. 2016 Mar;34(3):328-33. doi: 10.1038/nbt.3471. Epub 2016 Feb 1.

Therapeutic genome editing by combined viral and non-viral delivery of CRISPR system components in vivo.

Author information

  • 1David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
  • 2RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
  • 3Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
  • 4Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
  • 5Department of Molecular, Cell and Cancer Biology, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
  • 6Program in Bioinformatics and Integrative Biology, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
  • 7Department of Bioinformatics, School of Life Science and Technology, Tongji University, Shanghai, P.R. China.
  • 8Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
  • 9Gene Therapy Center, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
  • 10College of Pharmacy, the Ohio State University, Columbus, Ohio, USA.
  • 11Skolkovo Institute of Science and Technology, Skolkovo, Russia.
  • 12Department of Chemistry, M.V. Lomonosov Moscow State University, Leninskie Gory, Russia.
  • 13Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
  • 14Harvard-MIT Division of Health Sciences &Technology, Cambridge, Massachusetts, USA.
  • 15Institute of Medical Engineering and Science, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.

Abstract

The combination of Cas9, guide RNA and repair template DNA can induce precise gene editing and the correction of genetic diseases in adult mammals. However, clinical implementation of this technology requires safe and effective delivery of all of these components into the nuclei of the target tissue. Here, we combine lipid nanoparticle-mediated delivery of Cas9 mRNA with adeno-associated viruses encoding a sgRNA and a repair template to induce repair of a disease gene in adult animals. We applied our delivery strategy to a mouse model of human hereditary tyrosinemia and show that the treatment generated fumarylacetoacetate hydrolase (Fah)-positive hepatocytes by correcting the causative Fah-splicing mutation. Treatment rescued disease symptoms such as weight loss and liver damage. The efficiency of correction was >6% of hepatocytes after a single application, suggesting potential utility of Cas9-based therapeutic genome editing for a range of diseases.

PMID:
26829318
DOI:
10.1038/nbt.3471
[PubMed - indexed for MEDLINE]
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