Brief Communication

Genome editing with Cas9 in adult mice corrects a disease mutation and phenotype

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Abstract

We demonstrate CRISPR-Cas9–mediated correction of a Fah mutation in hepatocytes in a mouse model of the human disease hereditary tyrosinemia. Delivery of components of the CRISPR-Cas9 system by hydrodynamic injection resulted in initial expression of the wild-type Fah protein in 1/250 liver cells. Expansion of Fah-positive hepatocytes rescued the body weight loss phenotype. Our study indicates that CRISPR-Cas9–mediated genome editing is possible in adult animals and has potential for correction of human genetic diseases.

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Change history

  • Corrected online 31 March 2014

    In the version of this article initially published online, in the legend for Figure 1d, the scale bars are 100 μm and 20 μm, not “mm.” The error has been corrected for the print, PDF and HTML versions of this article.

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Acknowledgements

We thank I. Zhuang and W. Cai for technical assistance, F. Zhang for sharing pX330 CRISPR vectors, and D. Crowley and K. Cormier for histology. This work was supported in part by grants 2-PO1-CA42063 to P.A.S. and T.J. and core grant P30-CA14051 from the National Cancer Institute. This work was supported in part by National Institutes of Health (NIH) Grant R01-CA133404 and the Marie-D. & Pierre Casimir-Lambert Fund to P.A.S. T.J. is a Howard Hughes Investigator, the David H. Koch Professor of Biology and a Daniel K. Ludwig Scholar. H.Y. and S.C. are supported by 5-U54-CA151884-04 NIH Centers for Cancer Nanotechnology Excellence and the Harvard-MIT Center of Cancer Nanotechnology Excellence. W.X. is supported by grant 1K99CA169512. S.C. is a Damon Runyon Fellow (DRG-2117-12). The authors acknowledge the service of the late Sean Collier to the MIT community. We thank the Swanson Biotechnology Center for technical support.

Author information

Author notes

    • Hao Yin
    •  & Wen Xue

    These authors contributed equally to this work.

Affiliations

  1. David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.

    • Hao Yin
    • , Wen Xue
    • , Sidi Chen
    • , Roman L Bogorad
    • , Phillip A Sharp
    • , Tyler Jacks
    •  & Daniel G Anderson
  2. Oregon Stem Cell Center, Department of Pediatrics, Oregon Health and Science University, Portland, Oregon, USA.

    • Eric Benedetti
    •  & Markus Grompe
  3. Skolkovo Institute of Science and Technology, Skolkovo, Russian Federation.

    • Victor Koteliansky
  4. Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.

    • Phillip A Sharp
    •  & Tyler Jacks
  5. Howard Hughes Medical Institute, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.

    • Tyler Jacks
  6. Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.

    • Daniel G Anderson
  7. Harvard-MIT Division of Health Sciences & Technology, Cambridge, Massachusetts, USA.

    • Daniel G Anderson
  8. Institute of Medical Engineering and Science, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.

    • Daniel G Anderson

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Contributions

H.Y., W.X. and D.G.A. designed the study. H.Y., W.X., S.C., R.L.B. and E.B. performed experiments and analyzed data. M.G., V.K. and P.A.S. provided reagents and conceptual advice. H.Y., W.X., T.J. and D.G.A. wrote the manuscript.

Competing interests

D.G.A., H.Y., R.L.B., T.J. and W.X. have applied for patents on the subject matter of this paper.

Corresponding author

Correspondence to Daniel G Anderson.

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–8, Supplementary Discussion, Supplementary Methods and Supplementary Tables 1 and 2

Excel files

  1. 1.

    Supplementary Table 3

    Next-generation sequencing data for FAH2 treated mice.

  2. 2.

    Supplementary Table 4

    Next-generation sequencing data for off-target analysis of FAH2.