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Efficient DNA-free genome editing of bread wheat using CRISPR/Cas9 ribonucleoprotein complexes

Open access
Liang, Zhen, Chen, Kunling, Li, Tingdong, Zhang, Yi, Wang, Yanpeng, Zhao, Qian, Liu, Jinxing, Zhang, Huawei, Liu, Cuimin, Ran, Yidong, Gao, Caixia·2017·Nature Communications

Abstract

Substantial efforts are being made to optimize the CRISPR/Cas9 system for precision crop breeding. The avoidance of transgene integration and reduction of off-target mutations are the most important targets for optimization. Here, we describe an efficient genome editing method for bread wheat using CRISPR/Cas9 ribonucleoproteins (RNPs). Starting from RNP preparation, the whole protocol takes only seven to nine weeks, with four to five independent mutants produced from 100 immature wheat embryos. Deep sequencing reveals that the chance of off-target mutations in wheat cells is much lower in RNP mediated genome editing than in editing with CRISPR/Cas9 DNA. Consistent with this finding, no off-target mutations are detected in the mutant plants. Because no foreign DNA is used in CRISPR/Cas9 RNP mediated genome editing, the mutants obtained are completely transgene free. This method may be widely applicable for producing genome edited crop plants and has a good prospect of being commercialized.

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Metadata

Authors
Liang, Z., Chen, K., Li, T., Zhang, Y., Wang, Y., Zhao, Q., Liu, J., Zhang, H., Liu, C., Ran, Y., & Gao, C.
Venue
Nature Communications
Published
2017-01-18
Volume
8
Issue
1
Pages
14261-14261
ISSN
2041-1723 (ISSN-L)
OpenAlex
W2572900050
Cited by
955
References
2
FWCI
36.20
Type
journal article
Language
en
Open access
gold

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CoChat. (2026). Citation check: Efficient DNA-free genome editing of bread wheat using CRISPR/Cas9 ribonucleoprotein complexes. https://app.cochat.ai/papers/10.1038/ncomms14261
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Liang, Z., Chen, K., Li, T., Zhang, Y., Wang, Y., Zhao, Q., Liu, J., Zhang, H., Liu, C., Ran, Y., & Gao, C. (2017). Efficient DNA-free genome editing of bread wheat using CRISPR/Cas9 ribonucleoprotein complexes. Nature Communications. https://doi.org/10.1038/ncomms14261

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