CRISPR/Cas9 Based Cell-Type Specific Gene Knock-Out in <i>Arabidopsis</i> Roots
CRISPR/Cas9 (hereafter Cas9)-mediated gene knockout is one of the most important tools for studying gene function. However, many genes in plants play distinct roles in different cell types. Engineering the currently used Cas9 system to achieve cell-type-specific knockout of functional genes is usefu...
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| Format: | Article |
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MDPI AG
2023-06-01
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| Series: | Plants |
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| Online Access: | https://www.mdpi.com/2223-7747/12/12/2365 |
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| author | Meng Li Xufang Niu Shuang Li Shasha Fu Qianfang Li Meizhi Xu Chunhua Wang Shuang Wu |
| author_facet | Meng Li Xufang Niu Shuang Li Shasha Fu Qianfang Li Meizhi Xu Chunhua Wang Shuang Wu |
| author_sort | Meng Li |
| collection | DOAJ |
| description | CRISPR/Cas9 (hereafter Cas9)-mediated gene knockout is one of the most important tools for studying gene function. However, many genes in plants play distinct roles in different cell types. Engineering the currently used Cas9 system to achieve cell-type-specific knockout of functional genes is useful for addressing the cell-specific functions of genes. Here we employed the cell-specific promoters of the <i>WUSCHEL RELATED HOMEOBOX 5 </i>(<i>WOX5</i>), <i>CYCLIND6;1</i> (<i>CYCD6;1</i>), and <i>ENDODERMIS7</i> (<i>EN7</i>) genes to drive the Cas9 element, allowing tissue-specific targeting of the genes of interest. We designed the reporters to verify the tissue-specific gene knockout in vivo. Our observation of the developmental phenotypes provides strong evidence for the involvement of <i>SCARECROW</i> (<i>SCR</i>) and <i>GIBBERELLIC ACID INSENSITIVE</i> (<i>GAI</i>) in the development of quiescent center (QC) and endodermal cells. This system overcomes the limitations of traditional plant mutagenesis techniques, which often result in embryonic lethality or pleiotropic phenotypes. By allowing cell-type-specific manipulation, this system has great potential to help us better understand the spatiotemporal functions of genes during plant development. |
| first_indexed | 2024-03-11T02:00:51Z |
| format | Article |
| id | doaj.art-4537ea83bf6743cc931da817467233c6 |
| institution | Directory Open Access Journal |
| issn | 2223-7747 |
| language | English |
| last_indexed | 2024-03-11T02:00:51Z |
| publishDate | 2023-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Plants |
| spelling | doaj.art-4537ea83bf6743cc931da817467233c62023-11-18T12:11:07ZengMDPI AGPlants2223-77472023-06-011212236510.3390/plants12122365CRISPR/Cas9 Based Cell-Type Specific Gene Knock-Out in <i>Arabidopsis</i> RootsMeng Li0Xufang Niu1Shuang Li2Shasha Fu3Qianfang Li4Meizhi Xu5Chunhua Wang6Shuang Wu7College of Life Sciences and Horticultural Plant Biology Metabolomics Center, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCollege of Life Sciences and Horticultural Plant Biology Metabolomics Center, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCollege of Life Sciences and Horticultural Plant Biology Metabolomics Center, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCollege of Life Sciences and Horticultural Plant Biology Metabolomics Center, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCollege of Life Sciences and Horticultural Plant Biology Metabolomics Center, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCollege of Life Sciences and Horticultural Plant Biology Metabolomics Center, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCollege of Life Sciences and Horticultural Plant Biology Metabolomics Center, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCollege of Life Sciences and Horticultural Plant Biology Metabolomics Center, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCRISPR/Cas9 (hereafter Cas9)-mediated gene knockout is one of the most important tools for studying gene function. However, many genes in plants play distinct roles in different cell types. Engineering the currently used Cas9 system to achieve cell-type-specific knockout of functional genes is useful for addressing the cell-specific functions of genes. Here we employed the cell-specific promoters of the <i>WUSCHEL RELATED HOMEOBOX 5 </i>(<i>WOX5</i>), <i>CYCLIND6;1</i> (<i>CYCD6;1</i>), and <i>ENDODERMIS7</i> (<i>EN7</i>) genes to drive the Cas9 element, allowing tissue-specific targeting of the genes of interest. We designed the reporters to verify the tissue-specific gene knockout in vivo. Our observation of the developmental phenotypes provides strong evidence for the involvement of <i>SCARECROW</i> (<i>SCR</i>) and <i>GIBBERELLIC ACID INSENSITIVE</i> (<i>GAI</i>) in the development of quiescent center (QC) and endodermal cells. This system overcomes the limitations of traditional plant mutagenesis techniques, which often result in embryonic lethality or pleiotropic phenotypes. By allowing cell-type-specific manipulation, this system has great potential to help us better understand the spatiotemporal functions of genes during plant development.https://www.mdpi.com/2223-7747/12/12/2365CRISPR/Cas9genes functioncell-type-specific gene knock-outroot |
| spellingShingle | Meng Li Xufang Niu Shuang Li Shasha Fu Qianfang Li Meizhi Xu Chunhua Wang Shuang Wu CRISPR/Cas9 Based Cell-Type Specific Gene Knock-Out in <i>Arabidopsis</i> Roots Plants CRISPR/Cas9 genes function cell-type-specific gene knock-out root |
| title | CRISPR/Cas9 Based Cell-Type Specific Gene Knock-Out in <i>Arabidopsis</i> Roots |
| title_full | CRISPR/Cas9 Based Cell-Type Specific Gene Knock-Out in <i>Arabidopsis</i> Roots |
| title_fullStr | CRISPR/Cas9 Based Cell-Type Specific Gene Knock-Out in <i>Arabidopsis</i> Roots |
| title_full_unstemmed | CRISPR/Cas9 Based Cell-Type Specific Gene Knock-Out in <i>Arabidopsis</i> Roots |
| title_short | CRISPR/Cas9 Based Cell-Type Specific Gene Knock-Out in <i>Arabidopsis</i> Roots |
| title_sort | crispr cas9 based cell type specific gene knock out in i arabidopsis i roots |
| topic | CRISPR/Cas9 genes function cell-type-specific gene knock-out root |
| url | https://www.mdpi.com/2223-7747/12/12/2365 |
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