Brief Communication

Targeted base editing in rice and tomato using a CRISPR-Cas9 cytidine deaminase fusion

  • Nature Biotechnology volume 35, pages 441443 (2017)
  • doi:10.1038/nbt.3833
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Abstract

We applied a fusion of CRISPR-Cas9 and activation-induced cytidine deaminase (Target-AID) for point mutagenesis at genomic regions specified by single guide RNAs (sgRNAs) in two crop plants. In rice, we induced multiple herbicide-resistance point mutations by multiplexed editing using herbicide selection, while in tomato we generated marker-free plants with homozygous heritable DNA substitutions, demonstrating the feasibility of base editing for crop improvement.

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Change history

  • Corrected online 05 April 2017

    In the version of this article initially published, in the first paragraph after the abstract, a reference for Komor et al. was left out. The reference is now inserted in line 5 as “the base-editing1,18 construct”, replacing “with a construct.” The error has been corrected for the print, PDF and HTML versions of this article.

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Acknowledgements

Plant CRISPR vectors pCAS9-TPC and pChimera were kindly provided by F. Fauser, S. Schiml, and H. Puchta at Karlsruhe Institute of Technology via M. Endo and S. Toki. We thank Y. Iida, R. Ohta, Y. Ueke and M. Suzuki for technical assistance. This work was supported by the Cabinet Office, Government of Japan, Cross-ministerial Strategic Innovation Promotion Program (SIP), “Technologies for creating next-generation agriculture, forestry and fisheries” (funding agency: Bio-oriented Technology Research Advancement Institution, NARO) and partly supported by a Special Coordination Fund for Promoting Science and Technology, Creation of Innovative Centers for Advanced Interdisciplinary Research Areas (Innovative Bioproduction Kobe) from the Ministry of Education, Culture, Sports and Technology (MEXT) of Japan, by the Platform Project for Supporting in Drug Discovery and Life Science Research (Platform for Drug Discovery, Informatics, and Structural Life Science) from the Japan Agency for Medical Research and Development (AMED), by JSPS KAKENHI grant numbers 26119710 and 16K14654 and by the New Energy and Industrial Technology Development Organization (NEDO).

Author information

Author notes

    • Zenpei Shimatani
    • , Sachiko Kashojiya
    •  & Mariko Takayama

    These authors contributed equally to this work.

Affiliations

  1. Graduate School of Science, Technology and Innovation, Kobe University, Kobe, Japan.

    • Zenpei Shimatani
    • , Takayuki Arazoe
    • , Hisaki Ishii
    • , Hiroshi Teramura
    • , Keiji Nishida
    •  & Akihiko Kondo
  2. Faculty of Life and Environmental Sciences, University of Tsukuba, Gene Research Center, Tsukuba, Japan.

    • Sachiko Kashojiya
    • , Mariko Takayama
    • , Tsuyoshi Yamamoto
    • , Hiroki Komatsu
    • , Kenji Miura
    • , Hiroshi Ezura
    •  & Tohru Ariizumi
  3. Laboratory of Genetics and Breeding Science, Faculty of Agriculture, Meijo University, Nagoya, Japan.

    • Rie Terada
  4. Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, Kobe, Japan.

    • Akihiko Kondo

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Contributions

Z.S., H.I., H.T. and R.T. performed rice experiments. M.T., S.K., T.Y. and H.K. performed tomato experiments. K.N. and T. Arazoe performed NGS analysis. K.N. designed and constructed plasmids. Z.S., T. Ariizumi, R.T. and K.N. designed the experiments. Z.S., K.N., M.T., S.K. and T. Ariizumi, wrote the manuscript. K.N., T. Ariizumi, K.M., H.E. and A.K. supervised the project.

Competing interests

K.N., Z.S., T. Arazoe and A.K. have filed a patent related to the rice study as PCT/JP2016/085075. S.K., M.T., T.Y., K.M., T. Ariizumi and H.E. have filed a patent related to the tomato study as JP 2017-019921.

Corresponding authors

Correspondence to Hiroshi Ezura or Keiji Nishida or Tohru Ariizumi.

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Supplementary information

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  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–6

  2. 2.

    Supplementary Tables 1–8