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Nanoparticle delivery of CRISPR into the brain rescues a mouse model of fragile X syndrome from exaggerated repetitive behaviours

Nature Biomedical Engineeringvolume 2pages497507 (2018) | Download Citation


Technologies that can safely edit genes in the brains of adult animals may revolutionize the treatment of neurological diseases and the understanding of brain function. Here, we demonstrate that intracranial injection of CRISPR–Gold, a nonviral delivery vehicle for the CRISPR–Cas9 ribonucleoprotein, can edit genes in the brains of adult mice in multiple mouse models. CRISPR–Gold can deliver both Cas9 and Cpf1 ribonucleoproteins, and can edit all of the major cell types in the brain, including neurons, astrocytes and microglia, with undetectable levels of toxicity at the doses used. We also show that CRISPR–Gold designed to target the metabotropic glutamate receptor 5 (mGluR5) gene can efficiently reduce local mGluR5 levels in the striatum after an intracranial injection. The effect can also rescue mice from the exaggerated repetitive behaviours caused by fragile X syndrome, a common single-gene form of autism spectrum disorders. CRISPR–Gold may significantly accelerate the development of brain-targeted therapeutics and enable the rapid development of focal brain-knockout animal models.

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We thank J. Doudna for advice, B. Staahl for discussions and technical support, and H. Kim, A. Rao and K. Kataoka for technical support. We thank M. A. Bhat and members of the Bhat Lab for technical support. We thank M. West in the CIRM/QB3 Shared Stem Cell facility for technical support. This work was supported by the National Institutes of Health grant R01EB023776 to N.M, and by the National Science Foundation grant 1456862 to R.B.

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Author notes

  1. These authors contributed equally: Bumwhee Lee, Kunwoo Lee.


  1. The Department of Cellular and Integrative Physiology, University of Texas Health Science Center at San Antonio, San Antonio, TX, USA

    • Bumwhee Lee
    • , Shree Panda
    • , Rodrigo Gonzales-Rojas
    • , Vladislav Bugay
    • , Robert Brenner
    •  & Hye Young Lee
  2. GenEdit Inc., Berkeley, CA, USA

    • Kunwoo Lee
    • , Anthony Chong
    •  & Hyo Min Park
  3. Department of Bioengineering, University of California, Berkeley, Berkeley, CA, USA

    • Niren Murthy


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B.L., K.L., R.B., N.M. and H.Y.L. designed the research, and B.L., K.L., S.P., R.G.-R., A.C., H.M.P., V.B. and H.Y.L. performed the experiments and analyses. R.G.-R. and S.P. generated the videos. B.L., K.L., N.M. and H.Y.L. wrote the manuscript. H.Y.L. supervised the research. All authors discussed the results and commented on the manuscript.

Competing interests

K.L., H.M.P. and N.M. are co-founders of GenEdit Inc. The remaining authors declare no competing interests.

Corresponding authors

Correspondence to Niren Murthy or Hye Young Lee.

Supplementary information

  1. Supplementary Information

    Supplementary figures, tables and video captions.

  2. Reporting Summary

  3. Supplementary Video 1

    Marble-bury assay.

  4. Supplementary Video 2

    Empty-cage observations.

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