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Efficient C-to-T base editing in plants using a fusion of nCas9 and human APOBEC3A

Nature Biotechnology volume 36, pages 950953 (2018) | Download Citation

Abstract

Base editors (BEs) have been used to create C-to-T substitutions in various organisms. However, editing with rat APOBEC1-based BE3 is limited to a 5-nt sequence editing window and is inefficient in GC contexts. Here, we show that a base editor fusion protein composed of Cas9 nickase and human APOBEC3A (A3A-PBE) converts cytidine to thymidine efficiently in wheat, rice and potato with a 17-nucleotide editing window at all examined sites, independent of sequence context.

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Acknowledgements

We thank M. Andersson for technical support on potato protoplast regeneration and Y. Huo for technical support on flow cytometry. This work was supported by grants from the National Key Research and Development Program of China (2016YFD0101804), the National Natural Science Foundation of China (31788103 and 31420103912), the National Transgenic Science and Technology Program (2016ZX08010002, 2018ZX0800102B-001 and 2018ZX0801002B-002) and the Chinese Academy of Sciences (QYZDY-SSW-SMC030 and GJHZ1602).

Author information

Author notes

    • Yuan Zong
    •  & Qianna Song

    These authors contributed equally to this work.

Affiliations

  1. State Key Laboratory of Plant Cell and Chromosome Engineering, Center for Genome Editing, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, China.

    • Yuan Zong
    • , Qianna Song
    • , Chao Li
    • , Shuai Jin
    • , Dingbo Zhang
    • , Yanpeng Wang
    •  & Caixia Gao
  2. University of Chinese Academy of Sciences, Beijing, China.

    • Yuan Zong
    • , Qianna Song
    • , Chao Li
    • , Shuai Jin
    • , Dingbo Zhang
    •  & Caixia Gao
  3. State Key Laboratory of Plant Genomics, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.

    • Jin-Long Qiu

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Contributions

Y.Z. and C.G. designed the experiments; Y.Z., Q.S. and C.L. performed most of the experiments; D.Z. purified the protein. S.J. and Y.W. analyzed the results; C.G. supervised the project; Y.Z., J.-L.Q. and C.G. wrote the manuscript.

Competing interests

The authors have submitted a patent application (application no. 201810816603.7) based on the results reported in this paper.

Corresponding author

Correspondence to Caixia Gao.

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DOI

https://doi.org/10.1038/nbt.4261