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Conferring resistance to geminiviruses with the CRISPR–Cas prokaryotic immune system

Abstract

To reduce crop losses due to geminivirus infection, we targeted the bean yellow dwarf virus (BeYDV) genome for destruction with the CRISPR–Cas (clustered, regularly interspaced short palindromic repeats–CRISPR-associated proteins) system. Transient assays using BeYDV-based replicons revealed that CRISPR–Cas reagents introduced mutations within the viral genome and reduced virus copy number. Transgenic plants expressing CRISPR–Cas reagents and challenged with BeYDV had reduced virus load and symptoms, thereby demonstrating a novel strategy for engineering resistance to geminiviruses.

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Figure 1: Testing Cas9-sgRNA activity at targets within the BeYDV.
Figure 2: Restricting BeYDV infection with the CRISPR–Cas system.

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Acknowledgements

We thank K. Leffler for the help in generating the figures. We thank J. Gil for technical assistance with the qPCR experiment. This work was supported by a grant from the National Science Foundation (IOS-1339209).

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N.J.B. and A.W.H. designed the experiments, with input from D.F.V. N.J.B, A.W.H. and E.K. generated constructs. N.J.B., A.W.H. and E.K. performed the transient assays. E.K. provided technical support. R.C. analysed the NGS data. A.N.B. and D.M.B. helped design the infection experiments; E.K. and A.B. performed the experiments. N.J.B., A.W.H. and D.F.V. wrote the paper. D.F.V. supervised the entire project. All authors read and approved the final manuscript.

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Correspondence to Daniel F. Voytas.

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Competing interests

N.J.B., A.W.H. and D.F.V. are inventors on a patent application (WO2015048707A2) for the technology described in this work.

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Baltes, N., Hummel, A., Konecna, E. et al. Conferring resistance to geminiviruses with the CRISPR–Cas prokaryotic immune system. Nature Plants 1, 15145 (2015). https://doi.org/10.1038/nplants.2015.145

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