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Efficient and transgene-free genome editing in wheat through transient expression of CRISPR/Cas9 DNA or RNA

Author

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  • Yi Zhang

    (State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences
    University of Chinese Academy of Sciences)

  • Zhen Liang

    (State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences
    University of Chinese Academy of Sciences)

  • Yuan Zong

    (State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences
    University of Chinese Academy of Sciences)

  • Yanpeng Wang

    (State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences
    University of Chinese Academy of Sciences)

  • Jinxing Liu

    (State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences)

  • Kunling Chen

    (State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences)

  • Jin-Long Qiu

    (State Key Laboratory of Plant Genomics, Institute of Microbiology, Chinese Academy of Sciences)

  • Caixia Gao

    (State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences)

Abstract

Editing plant genomes is technically challenging in hard-to-transform plants and usually involves transgenic intermediates, which causes regulatory concerns. Here we report two simple and efficient genome-editing methods in which plants are regenerated from callus cells transiently expressing CRISPR/Cas9 introduced as DNA or RNA. This transient expression-based genome-editing system is highly efficient and specific for producing transgene-free and homozygous wheat mutants in the T0 generation. We demonstrate our protocol to edit genes in hexaploid bread wheat and tetraploid durum wheat, and show that we are able to generate mutants with no detectable transgenes. Our methods may be applicable to other plant species, thus offering the potential to accelerate basic and applied plant genome-engineering research.

Suggested Citation

  • Yi Zhang & Zhen Liang & Yuan Zong & Yanpeng Wang & Jinxing Liu & Kunling Chen & Jin-Long Qiu & Caixia Gao, 2016. "Efficient and transgene-free genome editing in wheat through transient expression of CRISPR/Cas9 DNA or RNA," Nature Communications, Nature, vol. 7(1), pages 1-8, November.
    • Handle: RePEc:nat:natcom:v:7:y:2016:i:1:d:10.1038_ncomms12617
    DOI: 10.1038/ncomms12617
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    Cited by:

    1. Bindu Paudel & Deepthi E. Kolady & David Just & Evert Van der Sluis, 2023. "Determinants of consumer acceptance of gene‐edited foods and its implications for innovators and policymakers," Agribusiness, John Wiley & Sons, Ltd., vol. 39(3), pages 623-645, July.
    2. Paudel, Bindu & Kolady, Deepthi Elizabeth & Just, David R. & Van Der Sluis, Evert, 2021. "Determinants of consumer acceptance of genetically modified and gene-edited foods: Market and policy implications," 2021 Annual Meeting, August 1-3, Austin, Texas 313905, Agricultural and Applied Economics Association.
    3. Connor Thorpe & Weifeng Luo & Qing Ji & Alan L. Eggenberger & Aline S. Chicowski & Weihui Xu & Ritinder Sandhu & Keunsub Lee & Steven A. Whitham & Yiping Qi & Kan Wang & Shan Jiang, 2025. "Enhancing biolistic plant transformation and genome editing with a flow guiding barrel," Nature Communications, Nature, vol. 16(1), pages 1-14, December.

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