Letter | Published:

Genome editing of upstream open reading frames enables translational control in plants

Nature Biotechnology | Download Citation

Abstract

Translational regulation by upstream open reading frames (uORFs) is becoming established as a general mechanism for controlling the amount of protein that is synthesized from downstream primary ORFs (pORFs)1,2,3,4,5. We found that genome editing of endogenous uORFs in plants enabled the modulation of translation of mRNAs from four pORFs that are involved in either development or antioxidant biosynthesis. A single-guide RNA that targeted the region harboring a uORF initiation codon can produce multiple mutations. Following uORF editing, we observed varying amounts of mRNA translation in four pORFs. Notably, editing the uORF of LsGGP2, which encodes a key enzyme in vitamin C biosynthesis in lettuce, not only increased oxidation stress tolerance, but also increased ascorbate content by 150%. These data indicate that editing plant uORFs provides a generalizable, efficient method for manipulating translation of mRNA that could be applied to dissect biological mechanisms and improve crops.

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Acknowledgements

We thank J.-L. Qui (Institute of Microbiology) for helpful discussion and insightful comments on the manuscript, Y. Wang and G. Wang for advice on the high-performance liquid chromatography assay, and Q. Xie (Institute of Genetics and Developmental Biology) for providing antibodies to BRI1 and PAG. 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, 31420103912, 31501376 and 31570369), and the Chinese Academy of Sciences (QYZDY-SSW-SMC030 and GJHZ1602).

Author information

Author notes

    • Huawei Zhang
    • , Xiaomin Si
    •  & Xiang Ji

    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.

    • Huawei Zhang
    • , Xiaomin Si
    • , Xiang Ji
    • , Jinxing Liu
    • , Kunling Chen
    • , Daowen Wang
    •  & Caixia Gao
  2. University of Chinese Academy of Sciences, Beijing, China.

    • Xiaomin Si
    • , Xiang Ji
    • , Rong Fan
    • , Daowen Wang
    •  & Caixia Gao
  3. Hebei Laboratory of Agricultural Water-Saving, Center for Agricultural Resources Research, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Shijiazhuang, China.

    • Rong Fan

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Contributions

H.Z., X.S., X.J. and C.G. designed the experiments. X.S., H.Z., X.J., R.F. and J.L. performed the experiments. X.J. and K.C. analyzed the results. C.G. supervised the project and C.G., D.W. and H.Z. wrote the manuscript.

Competing interests

The authors have submitted a patent application (application number 201710976945.0) 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.4202