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

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-str...

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Autores principales:	Sang-Tae Kim, Minkyung Choi, Su-Ji Bae, Jin-Soo Kim
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	CRISPR Cas9 guide RNA ACQOS Arabidopsis salt stress Biology (General) QH301-705.5 Chemistry QD1-999
Acceso en línea:	https://doaj.org/article/2bbdd92e8b604df882798025a1a15334
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spelling	oai:doaj.org-article:2bbdd92e8b604df882798025a1a153342021-11-11T16:51:44ZThe Functional Association of <i>ACQOS/VICTR</i> with Salt Stress Resistance in <i>Arabidopsis thaliana</i> Was Confirmed by CRISPR-Mediated Mutagenesis10.3390/ijms2221113891422-00671661-6596https://doaj.org/article/2bbdd92e8b604df882798025a1a153342021-10-01T00:00:00Zhttps://www.mdpi.com/1422-0067/22/21/11389https://doaj.org/toc/1661-6596https://doaj.org/toc/1422-0067Clustered 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 (<i>ACQOS</i>; <i>AT5G46520</i>) and neighboring nucleotide-binding leucine-rich repeats (<i>NLR</i>s; <i>AT5G46510</i>) is associated with osmotic tolerance. To investigate the role of <i>ACQOS</i> 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 <i>ACQOS</i> is directly associated with salt resistance, while the neighboring NLR is not. Here, we confirmed previous findings suggesting the involvement of <i>ACQOS</i> in salt tolerance and demonstrated the usefulness of CRISPR-mediated mutagenesis in validating the functions of genes in a single genetic background.Sang-Tae KimMinkyung ChoiSu-Ji BaeJin-Soo KimMDPI AGarticleCRISPRCas9guide RNAACQOSArabidopsissalt stressBiology (General)QH301-705.5ChemistryQD1-999ENInternational Journal of Molecular Sciences, Vol 22, Iss 11389, p 11389 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	CRISPR Cas9 guide RNA ACQOS Arabidopsis salt stress Biology (General) QH301-705.5 Chemistry QD1-999
spellingShingle	CRISPR Cas9 guide RNA ACQOS Arabidopsis salt stress Biology (General) QH301-705.5 Chemistry QD1-999 Sang-Tae Kim Minkyung Choi Su-Ji Bae Jin-Soo Kim The Functional Association of <i>ACQOS/VICTR</i> with Salt Stress Resistance in <i>Arabidopsis thaliana</i> Was Confirmed by CRISPR-Mediated Mutagenesis
description	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 (<i>ACQOS</i>; <i>AT5G46520</i>) and neighboring nucleotide-binding leucine-rich repeats (<i>NLR</i>s; <i>AT5G46510</i>) is associated with osmotic tolerance. To investigate the role of <i>ACQOS</i> 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 <i>ACQOS</i> is directly associated with salt resistance, while the neighboring NLR is not. Here, we confirmed previous findings suggesting the involvement of <i>ACQOS</i> in salt tolerance and demonstrated the usefulness of CRISPR-mediated mutagenesis in validating the functions of genes in a single genetic background.
format	article
author	Sang-Tae Kim Minkyung Choi Su-Ji Bae Jin-Soo Kim
author_facet	Sang-Tae Kim Minkyung Choi Su-Ji Bae Jin-Soo Kim
author_sort	Sang-Tae Kim
title	The Functional Association of <i>ACQOS/VICTR</i> with Salt Stress Resistance in <i>Arabidopsis thaliana</i> Was Confirmed by CRISPR-Mediated Mutagenesis
title_short	The Functional Association of <i>ACQOS/VICTR</i> with Salt Stress Resistance in <i>Arabidopsis thaliana</i> Was Confirmed by CRISPR-Mediated Mutagenesis
title_full	The Functional Association of <i>ACQOS/VICTR</i> with Salt Stress Resistance in <i>Arabidopsis thaliana</i> Was Confirmed by CRISPR-Mediated Mutagenesis
title_fullStr	The Functional Association of <i>ACQOS/VICTR</i> with Salt Stress Resistance in <i>Arabidopsis thaliana</i> Was Confirmed by CRISPR-Mediated Mutagenesis
title_full_unstemmed	The Functional Association of <i>ACQOS/VICTR</i> with Salt Stress Resistance in <i>Arabidopsis thaliana</i> Was Confirmed by CRISPR-Mediated Mutagenesis
title_sort	functional association of <i>acqos/victr</i> with salt stress resistance in <i>arabidopsis thaliana</i> was confirmed by crispr-mediated mutagenesis
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/2bbdd92e8b604df882798025a1a15334
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The Functional Association of <i>ACQOS/VICTR</i> with Salt Stress Resistance in <i>Arabidopsis thaliana</i> Was Confirmed by CRISPR-Mediated Mutagenesis

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