title: Therapeutic genome editing by combined viral and non-viral delivery of CRISPR system components in vivo.
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.
authors.affiliation: David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
authors.forename: Hao
authors.initials: H
authors.lastname: Yin
authors.affiliation: RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, Massachusetts, USA.|Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
authors.forename: Chun-Qing
authors.initials: CQ
authors.lastname: Song
authors.affiliation: David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.|Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
authors.forename: Joseph R
authors.initials: JR
authors.lastname: Dorkin
authors.affiliation: Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, Massachusetts, USA.|Department of Molecular, Cell and Cancer Biology, University of Massachusetts Medical School, Worcester, Massachusetts, USA.|Program in Bioinformatics and Integrative Biology, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
authors.forename: Lihua J
authors.initials: LJ
authors.lastname: Zhu
authors.affiliation: Department of Bioinformatics, School of Life Science and Technology, Tongji University, Shanghai, P.R. China.
authors.forename: Yingxiang
authors.identifier: http://orcid.org/0000-0003-0835-9280
authors.initials: Y
authors.lastname: Li
authors.affiliation: David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
authors.forename: Qiongqiong
authors.initials: Q
authors.lastname: Wu
authors.affiliation: RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
authors.forename: Angela
authors.initials: A
authors.lastname: Park
authors.affiliation: David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
authors.forename: Junghoon
authors.initials: J
authors.lastname: Yang
authors.affiliation: David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
authors.forename: Sneha
authors.initials: S
authors.lastname: Suresh
authors.affiliation: RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
authors.forename: Aizhan
authors.initials: A
authors.lastname: Bizhanova
authors.affiliation: Department of Molecular, Cell and Cancer Biology, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
authors.forename: Ankit
authors.initials: A
authors.lastname: Gupta
authors.affiliation: Department of Molecular, Cell and Cancer Biology, University of Massachusetts Medical School, Worcester, Massachusetts, USA.|Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
authors.forename: Mehmet F
authors.initials: MF
authors.lastname: Bolukbasi
authors.affiliation: David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
authors.forename: Stephen
authors.initials: S
authors.lastname: Walsh
authors.affiliation: David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
authors.forename: Roman L
authors.initials: RL
authors.lastname: Bogorad
authors.affiliation: Gene Therapy Center, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
authors.forename: Guangping
authors.initials: G
authors.lastname: Gao
authors.affiliation: Program in Bioinformatics and Integrative Biology, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
authors.forename: Zhiping
authors.initials: Z
authors.lastname: Weng
authors.affiliation: College of Pharmacy, the Ohio State University, Columbus, Ohio, USA.
authors.forename: Yizhou
authors.initials: Y
authors.lastname: Dong
authors.affiliation: Skolkovo Institute of Science and Technology, Skolkovo, Russia.|Department of Chemistry, M.V. Lomonosov Moscow State University, Leninskie Gory, Russia.
authors.forename: Victor
authors.initials: V
authors.lastname: Koteliansky
authors.affiliation: Department of Molecular, Cell and Cancer Biology, University of Massachusetts Medical School, Worcester, Massachusetts, USA.|Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
authors.forename: Scot A
authors.initials: SA
authors.lastname: Wolfe
authors.affiliation: David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.|Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.|Harvard-MIT Division of Health Sciences &Technology, Cambridge, Massachusetts, USA.|Institute of Medical Engineering and Science, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
authors.forename: Robert
authors.initials: R
authors.lastname: Langer
authors.affiliation: RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, Massachusetts, USA.|Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, Massachusetts, USA.|Department of Molecular, Cell and Cancer Biology, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
authors.forename: Wen
authors.initials: W
authors.lastname: Xue
authors.affiliation: David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.|Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.|Harvard-MIT Division of Health Sciences &Technology, Cambridge, Massachusetts, USA.|Institute of Medical Engineering and Science, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
authors.forename: Daniel G
authors.initials: DG
authors.lastname: Anderson
mesh_terms: D000818:Animals; D064113:CRISPR-Cas Systems / Q000235:genetics*; D004195:Disease Models, Animal; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D015894:Genome, Human; D006801:Humans; D008055:Lipids / Q000737:chemistry; D051379:Mice; D009154:Mutation; D053758:Nanoparticles / Q000737:chemistry; D017393:RNA Editing; D020176:Tyrosinemias / Q000235:genetics / Q000628:therapy*; D014780:Viruses / Q000235:genetics
chemical_list: D008055:Lipids
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