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CRISPR/Cas9-mediated viral interference in plants

Bibliography:

Zahir Ali, Aala Abulfaraj, Ali Idris, Shakila Ali, Manal Tashkandi and Magdy M. Mahfouz. CRISPR/Cas9-mediated viral interference in plants. Genome Biology 2015, 16:238  doi:10.1186/s13059-015-0799-6

Authors:

Zahir Ali, Aala Abulfaraj, Ali Idris, Shakila Ali, Manal Tashkandi and Magdy M. Mahfouz

Keywords:

Plant genome engineering; CRISPR/Cas9 system; Synthetic site-specific nucleases; Viral-mediated genome editing; Virus resistance; Tomato yellow leaf curl virus ; Tobacco rattle virus

Year:

2015

Abstract:

Background
The CRISPR/Cas9 system provides bacteria and archaea with molecular immunity against invading phages and conjugative plasmids. Recently, CRISPR/Cas9 has been used for targeted genome editing in diverse eukaryotic species.

Results
In this study, we investigate whether the CRISPR/Cas9 system could be used in plants to confer molecular immunity against DNA viruses. We deliver sgRNAs specific for coding and non-coding sequences of tomato yellow leaf curl virus (TYLCV) into Nicotiana benthamiana plants stably overexpressing the Cas9 endonuclease, and subsequently challenge these plants with TYLCV. Our data demonstrate that the CRISPR/Cas9 system targeted TYLCV for degradation and introduced mutations at the target sequences. All tested sgRNAs exhibit interference activity, but those targeting the stem-loop sequence within the TYLCV origin of replication in the intergenic region (IR) are the most effective. N. benthamiana plants expressing CRISPR/Cas9 exhibit delayed or reduced accumulation of viral DNA, abolishing or significantly attenuating symptoms of infection. Moreover, this system could simultaneously target multiple DNA viruses.

Conclusions
These data establish the efficacy of the CRISPR/Cas9 system for viral interference in plants, thereby extending the utility of this technology and opening the possibility of producing plants resistant to multiple viral infections.

ISSN:

ISSN: 1474-760X