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Adenine base editing in mouse embryos and an adult mouse model of Duchenne muscular dystrophy

Nature Biotechnology volume 36, pages 536539 (2018) | Download Citation

Abstract

Adenine base editors (ABEs) composed of an engineered adenine deaminase and the Streptococcus pyogenes Cas9 nickase enable adenine-to-guanine (A-to-G) single-nucleotide substitutions in a guide RNA (gRNA)-dependent manner. Here we demonstrate application of this technology in mouse embryos and adult mice. We also show that long gRNAs enable adenine editing at positions one or two bases upstream of the window that is accessible with standard single guide RNAs (sgRNAs). We introduced the Himalayan point mutation in the Tyr gene by microinjecting ABE mRNA and an extended gRNA into mouse embryos, obtaining Tyr mutant mice with an albino phenotype. Furthermore, we delivered the split ABE gene, using trans-splicing adeno-associated viral vectors, to muscle cells in a mouse model of Duchenne muscular dystrophy to correct a nonsense mutation in the Dmd gene, demonstrating the therapeutic potential of base editing in adult animals.

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Acknowledgements

This work was supported by the Institute for Basic Science (IBS-R021-D1 to J.-S.K).

Author information

Author notes

    • Seuk-Min Ryu
    • , Taeyoung Koo
    • , Kyoungmi Kim
    •  & Kayeong Lim

    These authors contributed equally to this work.

Affiliations

  1. Center for Genome Engineering, Institute for Basic Science, Seoul, Republic of Korea.

    • Seuk-Min Ryu
    • , Taeyoung Koo
    • , Kyoungmi Kim
    • , Kayeong Lim
    • , Gayoung Baek
    • , Sang-Tae Kim
    • , Heon Seok Kim
    • , Da-eun Kim
    • , Hyunji Lee
    • , Eugene Chung
    •  & Jin-Soo Kim
  2. Department of Chemistry, Seoul National University, Seoul, Republic of Korea.

    • Seuk-Min Ryu
    • , Kayeong Lim
    • , Heon Seok Kim
    • , Da-eun Kim
    • , Eugene Chung
    •  & Jin-Soo Kim
  3. Department of Biomedical Sciences and Department of Physiology, Korea University College of Medicine, Seoul, Republic of Korea.

    • Kyoungmi Kim

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Contributions

S.-M.R., T.K., K.K., K.L., and J.-S.K. designed the research. S.-M.R., T.K., K.K., K.L., G.B., S.-T.K., H.S.K., D.K., H.L., and E.C. performed the experiments. J.-S.K. supervised the research. All authors discussed the results and commented on the manuscript.

Competing interests

J.-S.K. and T.K. have filed a patent application based on this work. J.-S.K. is a co-founder of and holds stock in ToolGen, Inc.

Corresponding author

Correspondence to Jin-Soo Kim.

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DOI

https://doi.org/10.1038/nbt.4148

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