Therapeutic genome editing by combined viral and non-viral delivery of CRISPR system components in vivo
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
Biochemistry, Biophysics, and Structural Biology | Bioinformatics | Biotechnology | Computational Biology | Genetics | Genomics | Integrative Biology | Systems Biology
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.
DOI of Published Version
Nat Biotechnol. 2016 Mar;34(3):328-33. doi: 10.1038/nbt.3471. Epub 2016 Feb 1. Link to article on publisher's site
Yin, Hao; Song, Chun-Qing; Dorkin, Joseph R.; Zhu, Lihua Julie; Li, Yingxiang; Wu, Qiongqiong; Park, Angela I.; Yang, Junghoon; Suresh, Sneha; Bizhanova, Aizhan; Gupta, Ankit; Bolukbasi, Mehmet Fatih; Walsh, Stephen; Bogorad, Roman L.; Gao, Guangping; Weng, Zhiping; Dong, Yizhou; Koteliansky, Victor; Wolfe, Scot A.; Langer, Robert; Xue, Wen; and Anderson, Daniel G., "Therapeutic genome editing by combined viral and non-viral delivery of CRISPR system components in vivo" (2016). Program in Bioinformatics and Integrative Biology Publications and Presentations. 81.