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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Bhaya, D., Davison, M. & Barrangou, R. CRISPR-Cas systems in bacteria and archaea: versatile small RNAs for adaptive defense and regulation. Annu. Rev. Genet. 45, 273–297 (2011).
Hsu, P. D., Lander, E. S. & Zhang, F. Development and applications of CRISPR-Cas9 for genome engineering. Cell 157, 1262–1278 (2014).
Mansoor, S. et al. Geminivirus disease complexes: an emerging threat. Trends Plant Sci. 8, 128–134 (2003).
Moffat, A. S. Geminiviruses emerge as serious crop threat. Science 286, 1835 (1999).
Gutierrez, C. Geminivirus DNA replication. Cell. Mol. Life Sci. 56, 313–329 (1999).
Stenger, D. C. & McMahon, C. L. Genotypic diversity of beet curly top virus populations in the Western United States. Phytopathology 87, 737–744 (1997).
Zhang, Z. et al. BSCTV C2 attenuates the degradation of SAMDC1 to suppress DNA methylation-mediated gene silencing in Arabidopsis. Plant Cell 23, 273–288 (2011).
Chen, H. et al. Up-regulation of LSB1/GDU3 affects geminivirus infection by activating the salicylic acid pathway. Plant J. 62, 12–23 (2010).
Xing, H. L. et al. A CRISPR/Cas9 toolkit for multiplex genome editing in plants. BMC Plant Biol 14, 327 (2014).
Shan, Q. et al. Genome editing in rice and wheat using the CRISPR/Cas system. Nature Protoc. 9, 2395–2410 (2014).
Horsch, R. B. et al. A simple and general method for transferring genes into plants. Science 227, 1229–1231 (1985).
Clough, S. J. & Bent, A. F. Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J. 16, 735–743 (1998).
Wroblewski, T. et al. Optimization of Agrobacterium-mediated transient assays of gene expression in lettuce, tomato and Arabidopsis. Plant Biotechnol. J. 3, 259–273 (2005).
Sera, T. Inhibition of virus DNA replication by artificial zinc finger proteins. J. Virol. 79, 2614–2619 (2005).
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).
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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