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.
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Claims 7
Claim · model-written summary
The CRISPR/Cas9 system targeted TYLCV for degradation in Nicotiana benthamiana.
- Taxon
- Nicotiana benthamiana
Nicotiana benthamianae0 - Treatment
- CRISPR/Cas9 systeme1
- Interactor
- TYLCVTomato yellow leaf curl viruse1pathogen
- Outcome
- degradatione1
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“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”
e1“Our data demonstrate that the CRISPR/Cas9 system targeted TYLCV for degradation and introduced mutations at the target sequences”
Claim · model-written summary
The CRISPR/Cas9 system introduced mutations at the TYLCV target sequences.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae0
- Treatment
- CRISPR/Cas9 systeme1
- Interactor
- TYLCVTomato yellow leaf curl viruse1pathogen
- Outcome
- mutationse1
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“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”
e1“Our data demonstrate that the CRISPR/Cas9 system targeted TYLCV for degradation and introduced mutations at the target sequences”
Claim · model-written summary
All tested sgRNAs exhibit interference activity against TYLCV.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae0
- Treatment
- sgRNAse1
- Outcome
- interference activitye1
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“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”
e1“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”
Claim · model-written summary
sgRNAs targeting the stem-loop sequence within the TYLCV origin of replication are the most effective of the tested sgRNAs.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae0
- Treatment
- targeting the stem-loop sequence within the TYLCV origin of replicatione1
- Interactor
- TYLCVTomato yellow leaf curl viruse1pathogen
- Outcome
- most effectivee1
- Direction
- most effectivee1
- Comparator
- All tested sgRNAse1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“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”
e1“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”
Claim · model-written summary
Cas9-expressing N. benthamiana plants show delayed or reduced accumulation of viral DNA after challenge.
- Taxon
- N. benthamianaNicotiana benthamianae0
- Treatment
- expressing CRISPR/Cas9e0
- Outcome
- accumulation of viral DNAe0
- Direction
- delayed or reducede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“N. benthamiana plants expressing CRISPR/Cas9 exhibit delayed or reduced accumulation of viral DNA, abolishing or significantly attenuating symptoms of infection”
Claim · model-written summary
Cas9-expressing N. benthamiana plants show abolished or significantly attenuated symptoms of infection.
- Taxon
- N. benthamianaNicotiana benthamianae0
- Treatment
- expressing CRISPR/Cas9e0
- Outcome
- symptoms of infectione0
- Direction
- abolishing or significantly attenuatinge0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“N. benthamiana plants expressing CRISPR/Cas9 exhibit delayed or reduced accumulation of viral DNA, abolishing or significantly attenuating symptoms of infection”
Claim · model-written summary
The system could simultaneously target multiple DNA viruses.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae0
- Treatment
- this systeme1
- Outcome
- target multiple DNA virusese1
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“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”
e1“Moreover, this system could simultaneously target multiple DNA viruses”
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See all 143 papers →Claims are grouped by organism, treatment and what was measured or targeted. A paper counts as positive when any of its claims in the group is positive.