Title

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

UMMS Affiliation

RNA Therapeutics Institute; Program in Bioinformatics and Integrative Biology; Department of Molecular, Cell and Cancer Biology; Department of Biochemistry and Molecular Pharmacology; Gene Therapy Center

Date

3-2016

Document Type

Article

Disciplines

Biochemistry, Biophysics, and Structural Biology | Bioinformatics | Biotechnology | Computational Biology | Genetics | Genomics | Integrative Biology | Systems Biology

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.

Rights and Permissions

Citation: Nat Biotechnol. 2016 Mar;34(3):328-33. doi: 10.1038/nbt.3471. Epub 2016 Feb 1. Link to article on publisher's site

Related Resources

Link to Article in PubMed

PubMed ID

26829318