Expanding the Editing Window of Cytidine Base Editors With the Rad51 DNA-Binding Domain in Rice
Recently developed base editors provide a powerful tool for plant research and crop improvement. Although a number of different deaminases and Cas proteins have been used to improve base editors the editing efficiency, and editing window are still not optimal. Fusion of a non-sequence-specific singl...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2022-04-01
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| Series: | Frontiers in Plant Science |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fpls.2022.865848/full |
| _version_ | 1818048204478873600 |
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| author | Chunjie Wei Hao Liu Wenwen Wang Pengyu Luo Qiuling Chen Rou Li Chong Wang José Ramón Botella Hui Zhang |
| author_facet | Chunjie Wei Hao Liu Wenwen Wang Pengyu Luo Qiuling Chen Rou Li Chong Wang José Ramón Botella Hui Zhang |
| author_sort | Chunjie Wei |
| collection | DOAJ |
| description | Recently developed base editors provide a powerful tool for plant research and crop improvement. Although a number of different deaminases and Cas proteins have been used to improve base editors the editing efficiency, and editing window are still not optimal. Fusion of a non-sequence-specific single-stranded DNA-binding domain (DBD) from the human Rad51 protein between Cas9 nickase and the deaminase has been reported to dramatically increase the editing efficiency and expand the editing window of base editors in the mammalian cell lines and mouse embryos. We report the use of this strategy in rice, by fusing a rice codon-optimized human Rad51 DBD to the cytidine base editors AncBE4max, AncBE4max-NG, and evoFERNY. Our results show that the addition of Rad51 DBD did not increase editing efficiency in the major editing window but the editing range was expanded in all the three systems. Replacing the human Rad51 DBD with the rice Rad51 DBD homolog also expanded the editing window effectively. |
| first_indexed | 2024-12-10T10:17:58Z |
| format | Article |
| id | doaj.art-1f9fc24cf48847ca90cec6b81293da0a |
| institution | Directory Open Access Journal |
| issn | 1664-462X |
| language | English |
| last_indexed | 2024-12-10T10:17:58Z |
| publishDate | 2022-04-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Plant Science |
| spelling | doaj.art-1f9fc24cf48847ca90cec6b81293da0a2022-12-22T01:52:57ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2022-04-011310.3389/fpls.2022.865848865848Expanding the Editing Window of Cytidine Base Editors With the Rad51 DNA-Binding Domain in RiceChunjie Wei0Hao Liu1Wenwen Wang2Pengyu Luo3Qiuling Chen4Rou Li5Chong Wang6José Ramón Botella7Hui Zhang8Shanghai Key Laboratory of Plant Molecular Sciences, Development Center of Plant Germplasm Resources, College of Life Sciences, Shanghai Normal University, Shanghai, ChinaShanghai Key Laboratory of Plant Molecular Sciences, Development Center of Plant Germplasm Resources, College of Life Sciences, Shanghai Normal University, Shanghai, ChinaShanghai Key Laboratory of Plant Molecular Sciences, Development Center of Plant Germplasm Resources, College of Life Sciences, Shanghai Normal University, Shanghai, ChinaShanghai Key Laboratory of Plant Molecular Sciences, Development Center of Plant Germplasm Resources, College of Life Sciences, Shanghai Normal University, Shanghai, ChinaShanghai Key Laboratory of Plant Molecular Sciences, Development Center of Plant Germplasm Resources, College of Life Sciences, Shanghai Normal University, Shanghai, ChinaShanghai Key Laboratory of Plant Molecular Sciences, Development Center of Plant Germplasm Resources, College of Life Sciences, Shanghai Normal University, Shanghai, ChinaShanghai Key Laboratory of Plant Molecular Sciences, Development Center of Plant Germplasm Resources, College of Life Sciences, Shanghai Normal University, Shanghai, ChinaSchool of Agriculture and Food Sciences, University of Queensland, Brisbane, QLD, AustraliaShanghai Key Laboratory of Plant Molecular Sciences, Development Center of Plant Germplasm Resources, College of Life Sciences, Shanghai Normal University, Shanghai, ChinaRecently developed base editors provide a powerful tool for plant research and crop improvement. Although a number of different deaminases and Cas proteins have been used to improve base editors the editing efficiency, and editing window are still not optimal. Fusion of a non-sequence-specific single-stranded DNA-binding domain (DBD) from the human Rad51 protein between Cas9 nickase and the deaminase has been reported to dramatically increase the editing efficiency and expand the editing window of base editors in the mammalian cell lines and mouse embryos. We report the use of this strategy in rice, by fusing a rice codon-optimized human Rad51 DBD to the cytidine base editors AncBE4max, AncBE4max-NG, and evoFERNY. Our results show that the addition of Rad51 DBD did not increase editing efficiency in the major editing window but the editing range was expanded in all the three systems. Replacing the human Rad51 DBD with the rice Rad51 DBD homolog also expanded the editing window effectively.https://www.frontiersin.org/articles/10.3389/fpls.2022.865848/fullssDBDRad51 DBDCBEsediting windowrice |
| spellingShingle | Chunjie Wei Hao Liu Wenwen Wang Pengyu Luo Qiuling Chen Rou Li Chong Wang José Ramón Botella Hui Zhang Expanding the Editing Window of Cytidine Base Editors With the Rad51 DNA-Binding Domain in Rice Frontiers in Plant Science ssDBD Rad51 DBD CBEs editing window rice |
| title | Expanding the Editing Window of Cytidine Base Editors With the Rad51 DNA-Binding Domain in Rice |
| title_full | Expanding the Editing Window of Cytidine Base Editors With the Rad51 DNA-Binding Domain in Rice |
| title_fullStr | Expanding the Editing Window of Cytidine Base Editors With the Rad51 DNA-Binding Domain in Rice |
| title_full_unstemmed | Expanding the Editing Window of Cytidine Base Editors With the Rad51 DNA-Binding Domain in Rice |
| title_short | Expanding the Editing Window of Cytidine Base Editors With the Rad51 DNA-Binding Domain in Rice |
| title_sort | expanding the editing window of cytidine base editors with the rad51 dna binding domain in rice |
| topic | ssDBD Rad51 DBD CBEs editing window rice |
| url | https://www.frontiersin.org/articles/10.3389/fpls.2022.865848/full |
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