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The Functional Association of ACQOS/VICTR with Salt Stress Resistance in Arabidopsis thaliana Was Confirmed by CRISPR-Mediated Mutagenesis

1
Department of Medical & Biological Sciences, The Catholic University of Korea, Bucheon 14662, Korea
2
Center for Genome Engineering, Institute for Basic Science, Daejeon 34126, Korea
*
Author to whom correspondence should be addressed.
Academic Editors: Ki-Hong Jung and Jae-Yean Kim
Int. J. Mol. Sci. 2021, 22(21), 11389; https://doi.org/10.3390/ijms222111389
Received: 25 August 2021 / Revised: 18 October 2021 / Accepted: 19 October 2021 / Published: 21 October 2021
Clustered regularly interspaced palindromic repeat (CRISPR)-mediated mutagenesis has become an important tool in plant research, enabling the characterization of genes via gene knock-out. CRISPR genome editing tools can be applied to generate multi-gene knockout lines. Typically, multiple single-stranded, single guide RNAs (gRNAs) must be expressed in an organism to target multiple genes simultaneously; however, a single gRNA can target multiple genes if the target genes share similar sequences. A gene cluster comprising ACQUIRED OSMOTOLERANCE (ACQOS; AT5G46520) and neighboring nucleotide-binding leucine-rich repeats (NLRs; AT5G46510) is associated with osmotic tolerance. To investigate the role of ACQOS and the tandemly arranged NLR in osmotic tolerance, we introduced small insertion/deletion mutations into two target genes using a single gRNA and obtained transformant plant lines with three different combinations of mutant alleles. We then tested our mutant lines for osmotic tolerance after a salt-stress acclimation period by determining the chlorophyll contents of the mutant seedlings. Our results strongly suggest that ACQOS is directly associated with salt resistance, while the neighboring NLR is not. Here, we confirmed previous findings suggesting the involvement of ACQOS in salt tolerance and demonstrated the usefulness of CRISPR-mediated mutagenesis in validating the functions of genes in a single genetic background. View Full-Text
Keywords: CRISPR; Cas9; guide RNA; ACQOS; Arabidopsis; salt stress; chlorophyll CRISPR; Cas9; guide RNA; ACQOS; Arabidopsis; salt stress; chlorophyll
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MDPI and ACS Style

Kim, S.-T.; Choi, M.; Bae, S.-J.; Kim, J.-S. The Functional Association of ACQOS/VICTR with Salt Stress Resistance in Arabidopsis thaliana Was Confirmed by CRISPR-Mediated Mutagenesis. Int. J. Mol. Sci. 2021, 22, 11389. https://doi.org/10.3390/ijms222111389

AMA Style

Kim S-T, Choi M, Bae S-J, Kim J-S. The Functional Association of ACQOS/VICTR with Salt Stress Resistance in Arabidopsis thaliana Was Confirmed by CRISPR-Mediated Mutagenesis. International Journal of Molecular Sciences. 2021; 22(21):11389. https://doi.org/10.3390/ijms222111389

Chicago/Turabian Style

Kim, Sang-Tae, Minkyung Choi, Su-Ji Bae, and Jin-Soo Kim. 2021. "The Functional Association of ACQOS/VICTR with Salt Stress Resistance in Arabidopsis thaliana Was Confirmed by CRISPR-Mediated Mutagenesis" International Journal of Molecular Sciences 22, no. 21: 11389. https://doi.org/10.3390/ijms222111389

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