Brief Communication | Published:

Precision genome engineering through adenine base editing in plants

Nature Plantsvolume 4pages427431 (2018) | Download Citation

Abstract

The recent development of adenine base editors (ABEs) has enabled efficient and precise A-to-G base conversions in higher eukaryotic cells. Here, we show that plant-compatible ABE systems can be successfully applied to protoplasts of Arabidopsis thaliana and Brassica napus through transient transfection, and to individual plants through Agrobacterium-mediated transformation to obtain organisms with desired phenotypes. Targeted, precise A-to-G substitutions generated a single amino acid change in the FT protein or mis-splicing of the PDS3 RNA transcript, and we could thereby obtain transgenic plants with late-flowering and albino phenotypes, respectively. Our results provide ‘proof of concept’ for in planta ABE applications that can lead to induced neo-functionalization or altered mRNA splicing, opening up new avenues for plant genome engineering and biotechnology.

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Change history

  • 23 August 2018

    In Supplementary Fig. 1b originally published with this Brief Communication, the DNA sequence of nickase Cas9 was incorrect; this has now been amended.

References

  1. 1.

    Yin, K., Gao, C. & Qiu, J.-L. Nat. Plants 3, 17107 (2017).

  2. 2.

    Woo, J. W. et al. Nat. Biotechnol. 33, 1162–1164 (2015).

  3. 3.

    Alonso-Blanco, C. et al. Plant Cell 21, 1877–1896 (2009).

  4. 4.

    Slade, A. J., Fuerstenberg, S. I., Loeffler, D., Steine, M. N. & Facciotti, D. Nat. Biotechnol. 23, 75–81 (2005).

  5. 5.

    Komor, A. C., Kim, Y. B., Packer, M. S., Zuris, J. A. & Liu, D. R. Nature 533, 420–424 (2016).

  6. 6.

    Nishida, K. et al. Science 353, 1248–1257 (2016).

  7. 7.

    Zong, Y. et al. Nat. Biotechnol. 35, 438–440 (2017).

  8. 8.

    Shimatani, Z. et al. Nat. Biotechnol. 35, 441–443 (2017).

  9. 9.

    Gaudelli, N. M. et al. Nature 551, 464–471 (2017).

  10. 10.

    Bae, S., Park, J. & Kim, J. S. Bioinformatics 30, 1473–1475 (2014).

  11. 11.

    Kim, D. et al. Nat. Methods 12, 237–243 (2015).

  12. 12.

    Yan, L. et al. Mol. Plant 8, 1820–1823 (2015).

  13. 13.

    Tsutsui, H. & Higashiyama, T. Plant Cell Physiol. 58, 46–56 (2017).

  14. 14.

    Hanzawa, Y., Money, T. & Bradley, D. Proc. Natl Acad. Sci. USA 102, 7748–7753 (2005).

  15. 15.

    Qin, G. et al. Cell Res. 17, 471–482 (2007).

  16. 16.

    Hua, K., Tao, X., Yuan, F., Wang, D. & Zhu, J.-K. Mol. Plant 11, 627–630 (2018).

  17. 17.

    Yan, F. et al. Mol. Plant 11, 631–634 (2018).

  18. 18.

    Kim, H. et al. J. Integr. Plant Biol. 58, 705–712 (2016).

  19. 19.

    Zhang, X., Henriques, R., Lin, S. S., Niu, Q. W. & Chua, N. H. Nat. Protoc. 1, 641–646 (2006).

  20. 20.

    Park, J., Lim, K., Kim, J.-S. & Bae, S. Bioinformatics 33, 286–288 (2017).

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Acknowledgements

We thank all members of the Center for Genome Engineering for their support. We thank J. Suh for comments on the manuscript and technical assistance. This work was supported by the research fund from the Institute for Basic Science, Republic of Korea (IBS-R021-D1).

Author information

Author notes

  1. These authors contributed equally: Beum-Chang Kang, Jae-Young Yun.

Affiliations

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

    • Beum-Chang Kang
    • , Jae-Young Yun
    • , Sang-Tae Kim
    • , YouJin Shin
    • , Jahee Ryu
    • , Minkyung Choi
    • , Je Wook Woo
    •  & Jin-Soo Kim
  2. IBS School, University of Science and Technology, Daejeon, Republic of Korea

    • Beum-Chang Kang
    •  & Jin-Soo Kim
  3. Department of Chemistry, Seoul National University, Seoul, Republic of Korea

    • Jin-Soo Kim

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Contributions

J.-Y.Y., B.-C.K., S.-T.K. and J.W.W. designed the experiments. B.-C.K., J.-Y.Y., J.R., Y.S. and M.C. generated all the constructs. B.-C.K. performed the transient assay in protoplasts. J.-Y.Y., Y.S. and J.R. generated the transgenic Arabidopsis and analysed the plants. J.-Y.Y., S.-T.K., B.-C.K. and J.-S.K. wrote the manuscript with the help of all other authors. J.-S.K. supervised the project.

Competing interests

J.-S.K. is a co-founder of and holds stocks in ToolGen, Inc. All other authors declare no competing interests.

Corresponding author

Correspondence to Jin-Soo Kim.

Supplementary information

  1. Supplementary Information

    Supplementary Figures 1–8, Supplementary Table 1

  2. Reporting Summary

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DOI

https://doi.org/10.1038/s41477-018-0178-x

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