Abstract
Gene targeting (GT) is a useful technology for accurate genome engineering in plants. A reproducible approach based on a positive-negative selection system using hygromycin resistance and the diphtheria toxin A subunit gene as positive and negative selection markers, respectively, is now available. However, to date, this selection system has been applied exclusively in rice (Oryza sativa). To establish a universally applicable positive-negative GT system in plants, we designed a selection system using a combination of neomycin phosphotransferaseII (nptII) and an antisense nptII construct. The concomitant transcription of both sense and antisense nptII suppresses significantly the level of expression of the sense nptII gene, and transgenic calli and plants become sensitive to the antibiotic geneticin. In addition, we were able to utilize the sense nptII gene as a positive selection marker and the antisense nptII construct as a negative selection marker for knockout of the endogenous rice genes Waxy and 33-kD globulin through GT, although negative selection with this system is relatively less efficient compared with diphtheria toxin A subunit. The approach developed here, with some additional improvements, could be applied as a universal selection system for the enrichment of GT cells in several plant species.
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Claims 5
Claim · model-written summary
Sense and antisense nptII transcription significantly suppressed sense nptII expression.
- Taxon
- rice
Oryza sativae1 - Treatment
- concomitant transcription of both sense and antisense nptIIe0
- Gene
- nptIIe0
- Outcome
- level of expression of the sense nptII genee0
- Direction
- suppressese0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The concomitant transcription of both sense and antisense nptII suppresses significantly the level of expression of the sense nptII gene”
e1“rice (Oryza sativa)”
Claim · model-written summary
Transgenic calli and plants became sensitive to geneticin.
- Taxon
- riceOryza sativae1
- Organ / stage
- calli and plantse0
- Outcome
- sensitive to the antibiotic geneticine0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“transgenic calli and plants become sensitive to the antibiotic geneticin.”
e1“rice (Oryza sativa)”
Claim · model-written summary
The selection system supported knockout of rice Waxy.
- Taxon
- riceOryza sativae0
- Gene
- Waxye0
- Outcome
- knockout of the endogenous rice genes Waxye0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“positive selection marker and the antisense nptII construct as a negative selection marker for knockout of the endogenous rice genes Waxy and 33-kD globulin through GT”
Claim · model-written summary
The selection system supported knockout of rice 33-kD globulin.
- Taxon
- riceOryza sativae0
- Gene
- 33-kD globuline0
- Outcome
- knockout of the endogenous rice genes Waxy and 33-kD globuline0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“knockout of the endogenous rice genes Waxy and 33-kD globulin through GT”
Claim · model-written summary
Negative selection was less efficient than with diphtheria toxin A subunit.
- Taxon
- riceOryza sativae1
- Outcome
- negative selection with this systeme0
- Direction
- less efficiente0
- Comparator
- diphtheria toxin A subunite0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“negative selection with this system is relatively less efficient compared with diphtheria toxin A subunit.”
e1“rice (Oryza sativa)”
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neomycin · molecular: expression · Oryza sativa (rice)Positive and negative results
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- negative
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- negative
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See all 98 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.