Abstract
Abstract Drought episodes decrease plant growth and productivity, which in turn cause high economic losses. Plants naturally sense and respond to water stress by activating specific signalling pathways leading to physiological and developmental adaptations. Genetically engineering genes that belong to these pathways might improve the drought tolerance of plants. The abscisic acid (ABA)-responsive element binding protein 1/ABRE binding factor (AREB1/ABF2) is a key positive regulator of the drought stress response. We investigated whether the CRISPR activation (CRISPRa) system that targets AREB1 might contribute to improve drought stress tolerance in Arabidopsis. Arabidopsis histone acetyltransferase 1 (AtHAT1) promotes gene expression activation by switching chromatin to a relaxed state. Stable transgenic plants expressing chimeric dCas9 HAT were first generated. Then, we showed that the CRISPRa dCas9 HAT mechanism increased the promoter activity controlling the β-glucuronidase ( GUS ) reporter gene. To activate the endogenous promoter of AREB1 , the CRISPRa dCas9 HAT system was set up, and resultant plants showed a dwarf phenotype. Our qRT-PCR experiments indicated that both AREB1 and RD29A , a gene positively regulated by AREB1, exhibited higher gene expression than the control plants. The plants generated here showed higher chlorophyll content and faster stomatal aperture under water deficit, in addition to a better survival rate after drought stress. Altogether, we report that CRISPRa dCas9 HAT is a valuable biotechnological tool to improve drought stress tolerance through the positive regulation of AREB1.
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Claims 8
Claim · model-written summary
CRISPRa dCas9 HAT increased GUS reporter promoter activity.
- Taxon
- Arabidopsis
Arabidopsise0 - Treatment
- CRISPRa dCas9 HATe1
- Gene
- β-glucuronidase ( GUS ) reporter genee1
- Outcome
- promoter activitye1
- Direction
- increasede1
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Arabidopsis histone acetyltransferase 1 (AtHAT1)”
e1“the CRISPRa dCas9 HAT mechanism increased the promoter activity controlling the β-glucuronidase ( GUS ) reporter gene”
Claim · model-written summary
Plants with endogenous AREB1 promoter activation had a dwarf phenotype.
- Taxon
- ArabidopsisArabidopsise0
- Gene
- AREB1e1
- Outcome
- dwarf phenotypee1
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Arabidopsis histone acetyltransferase 1 (AtHAT1)”
e1“To activate the endogenous promoter of AREB1 , the CRISPRa dCas9 HAT system was set up, and resultant plants showed a dwarf phenotype.”
Claim · model-written summary
AREB1 expression was higher than in control plants.
- Taxon
- ArabidopsisArabidopsise0
- Treatment
- CRISPRa dCas9 HATe2
- Gene
- AREB1e1
- Outcome
- gene expressione1
- Direction
- highere1
- Comparator
- control plantse1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Arabidopsis histone acetyltransferase 1 (AtHAT1)”
e1“both AREB1 and RD29A , a gene positively regulated by AREB1, exhibited higher gene expression than the control plants”
e2“the CRISPRa dCas9 HAT system was set up”
Claim · model-written summary
RD29A expression was higher than in control plants.
- Taxon
- ArabidopsisArabidopsise0
- Treatment
- CRISPRa dCas9 HATe2
- Gene
- RD29Ae1
- Outcome
- gene expressione1
- Direction
- highere1
- Comparator
- control plantse1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Arabidopsis histone acetyltransferase 1 (AtHAT1)”
e1“both AREB1 and RD29A , a gene positively regulated by AREB1, exhibited higher gene expression than the control plants”
e2“the CRISPRa dCas9 HAT system was set up”
Claim · model-written summary
Generated plants had higher chlorophyll content under water deficit.
- Taxon
- ArabidopsisArabidopsise0
- Treatment
- CRISPRa dCas9 HATe2
- Outcome
- chlorophyll contente1
- Direction
- highere1
- Qualifier
- under water deficite1
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Arabidopsis histone acetyltransferase 1 (AtHAT1)”
e1“The plants generated here showed higher chlorophyll content and faster stomatal aperture under water deficit”
e2“the CRISPRa dCas9 HAT system was set up”
Claim · model-written summary
Generated plants had faster stomatal aperture under water deficit.
- Taxon
- ArabidopsisArabidopsise0
- Treatment
- CRISPRa dCas9 HATe2
- Outcome
- stomatal aperturee1
- Direction
- fastere1
- Qualifier
- under water deficite1
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Arabidopsis histone acetyltransferase 1 (AtHAT1)”
e1“The plants generated here showed higher chlorophyll content and faster stomatal aperture under water deficit”
e2“the CRISPRa dCas9 HAT system was set up”
Claim · model-written summary
Generated plants had better survival after drought stress.
- Taxon
- ArabidopsisArabidopsise0
- Treatment
- CRISPRa dCas9 HATe2
- Outcome
- survival ratee1
- Direction
- bettere1
- Qualifier
- after drought stresse1
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Arabidopsis histone acetyltransferase 1 (AtHAT1)”
e1“a better survival rate after drought stress”
e2“the CRISPRa dCas9 HAT system was set up”
Claim · model-written summary
Authors present CRISPRa dCas9 HAT as a tool to improve drought tolerance through positive AREB1 regulation.
- Taxon
- ArabidopsisArabidopsise0
- Gene
- AREB1e1
- Outcome
- improve drought stress tolerancee1
Not reportedComparatorValue
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Schema note from the model: Tool value is asserted without numeric result or comparator.
Verbatim
e0“Arabidopsis histone acetyltransferase 1 (AtHAT1)”
e1“CRISPRa dCas9 HAT is a valuable biotechnological tool to improve drought stress tolerance through the positive regulation of AREB1”
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