Abstract
Genome editing using sequence-specific nucleases (SSNs) is rapidly being developed for genetic engineering in crop species. The utilization of zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and clustered regularly interspaced short palindromic repeats/CRISPR-associated systems (CRISPR/Cas) for inducing double-strand breaks facilitates targeting of virtually any sequence for modification. Targeted mutagenesis via non-homologous end-joining (NHEJ) has been demonstrated extensively as being the preferred DNA repair pathway in plants. However, gene targeting via homologous recombination (HR) remains more elusive but could be a powerful tool for directed DNA repair. To overcome barriers associated with gene targeting, a geminivirus replicon (GVR) was used to deliver SSNs targeting the potato ACETOLACTATE SYNTHASE1 (ALS1) gene and repair templates designed to incorporate herbicide-inhibiting point mutations within the ALS1 locus. Transformed events modified with GVRs held point mutations that were capable of supporting a reduced herbicide susceptibility phenotype, while events transformed with conventional T-DNAs held no detectable mutations and were similar to wild-type. Regeneration of transformed events improved detection of point mutations that supported a stronger reduced herbicide susceptibility phenotype. These results demonstrate the use of geminiviruses for delivering genome editing reagents in plant species, and a novel approach to gene targeting in a vegetatively propagated species.
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Claims 4
Claim · model-written summary
Potato events transformed with geminivirus replicons (GVRs) held point mutations capable of supporting reduced herbicide susceptibility, compared with conventional T-DNA transformation.
- Taxon
- potato
Solanum tuberosume2 - Treatment
- a geminivirus replicon (GVR)e2
- Gene
- ACETOLACTATE SYNTHASE1 (ALS1)e2
- Outcome
- herbicide susceptibilitye1
- Direction
- reducede1
- Comparator
- events transformed with conventional T-DNAse1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e1“Transformed events modified with GVRs held point mutations that were capable of supporting a reduced herbicide susceptibility phenotype, while events transformed with conventional T-DNAs held no detectable mutations and were similar to wild-type”
e2“a geminivirus replicon (GVR) was used to deliver SSNs targeting the potato ACETOLACTATE SYNTHASE1 (ALS1) gene”
Claim · model-written summary
Potato events transformed with conventional T-DNAs held no detectable mutations and were similar to wild-type.
- Taxon
- potatoSolanum tuberosume2
- Treatment
- conventional T-DNAse3
- Outcome
- detectable mutationse3
- Qualifier
- similar to wild-typee3
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e2“a geminivirus replicon (GVR) was used to deliver SSNs targeting the potato ACETOLACTATE SYNTHASE1 (ALS1) gene”
e3“events transformed with conventional T-DNAs held no detectable mutations and were similar to wild-type”
Claim · model-written summary
Regeneration of transformed potato events improved detection of point mutations.
- Taxon
- potatoSolanum tuberosume2
- Treatment
- Regeneration of transformed eventse4
- Outcome
- detection of point mutationse4
- Direction
- improvede4
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e2“a geminivirus replicon (GVR) was used to deliver SSNs targeting the potato ACETOLACTATE SYNTHASE1 (ALS1) gene”
e4“Regeneration of transformed events improved detection of point mutations that supported a stronger reduced herbicide susceptibility phenotype”
Claim · model-written summary
Regeneration of transformed potato events supported a stronger reduced herbicide susceptibility phenotype.
- Taxon
- potatoSolanum tuberosume2
- Treatment
- Regeneration of transformed eventse4
- Outcome
- herbicide susceptibility phenotypee4
- Direction
- stronger reducede4
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e2“a geminivirus replicon (GVR) was used to deliver SSNs targeting the potato ACETOLACTATE SYNTHASE1 (ALS1) gene”
e4“Regeneration of transformed events improved detection of point mutations that supported a stronger reduced herbicide susceptibility phenotype”
The same questions in other papers
transgenic · disease · Solanum tuberosum (potato)Positive and negative results
14 papers in the same direction as this paper · 3 in the opposite direction
- negative
- negative
- no effect
14 papers more in this group
See all 18 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.