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Establishing a CRISPR–Cas-like immune system conferring DNA virus resistance in plants

Abstract

CRISPR–Cas (clustered, regularly interspaced short palindromic repeats–CRISPR-associated proteins) is an adaptive immune system in many archaea and bacteria that cleaves foreign DNA on the basis of sequence complementarity. Here, using the geminivirus, beet severe curly top virus (BSCTV), transient assays performed in Nicotiana benthamiana demonstrate that the sgRNA–Cas9 constructs inhibit virus accumulation and introduce mutations at the target sequences. Further, transgenic Arabidopsis and N. benthamiana plants overexpressing sgRNA–Cas9 are highly resistant to virus infection.

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Figure 1: Screening viral target sites and direct cutting of virus in N. benthamiana.
Figure 2: Transgenic N. benthamiana and Arabidopsis plants develop resistance to BSCTV.

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Acknowledgements

The authors thank Q. Xie (Institute of Genetics and Developmental Biology, CAS) for providing pCambia-BSCTV vector. Q. Chen (China Agricultural University) for providing pHSN401 vector and Q. Shen (Institute of Genetics and Developmental Biology, CAS) for providing N. benthamiana plants. This work was supported by grants from the National Natural Science Foundation of China (31420103912 and 31271795) and the Ministry of Agriculture of China (2014ZX0801003B).

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H.W.Z., C.X.G. and X.J. designed the experiments; X.J. performed most of the experiments; H.W.Z. and Y.Z. performed the virus-cutting experiments and mutation analysis; all authors analysed the data; C.X.G., X.J. and H.W.Z. wrote the manuscript. X.J. and Y.P.W. generated the figures. All authors approved the manuscript.

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Correspondence to Caixia Gao.

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X.J., H.W.Z. and C.X.G. filed a patent application in China (priority filing with serial number 201510107492.9).

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Ji, X., Zhang, H., Zhang, Y. et al. Establishing a CRISPR–Cas-like immune system conferring DNA virus resistance in plants. Nature Plants 1, 15144 (2015). https://doi.org/10.1038/nplants.2015.144

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