Application of CRISPR/CasΦ2 System for Genome Editing in Plants
CRISPR/Cas system has developed a new technology to modify target genes. In this study, CasΦ2 is a newly Cas protein that we used for genome modification in Arabidopsis and tobacco. <i>PDS</i> and <i>BRI1</i> of marker genes were chosen for targeting. CasΦ2 has the function t...
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| Format: | Article |
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MDPI AG
2022-05-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/10/5755 |
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| author | Qinan Cai Dongmei Guo Yujun Cao Yuan Li Rui Ma Wenping Liu |
| author_facet | Qinan Cai Dongmei Guo Yujun Cao Yuan Li Rui Ma Wenping Liu |
| author_sort | Qinan Cai |
| collection | DOAJ |
| description | CRISPR/Cas system has developed a new technology to modify target genes. In this study, CasΦ2 is a newly Cas protein that we used for genome modification in Arabidopsis and tobacco. <i>PDS</i> and <i>BRI1</i> of marker genes were chosen for targeting. CasΦ2 has the function to cleave pre-crRNA. In the presence of 10 mM Mg<sup>2+</sup> irons concentration, sgRNA3 type guided CasΦ2 to edit target gene and generate mutation, and a mutant seedling of <i>AtBRI1</i> gene with an expected male sterile phenotype was obtained. In the process of tobacco transformation, the gene editing activity of CasΦ2 can be activated by 100 nM Mg<sup>2+</sup> irons concentration, and sgRNA1 type guided CasΦ2 to edit target gene. Mutant seedlings of <i>NtPDS</i> gene with an expected albino were obtained. The results indicate that CasΦ2 can effectively edit target genes under the guidance of different sgRNA type in the presence of Mg<sup>2+</sup> ions. Together, our results verify that the CRISPR/CasΦ2 system is an effective and precise tool for genome editing in plants. |
| first_indexed | 2024-03-10T03:43:02Z |
| format | Article |
| id | doaj.art-f870682d0d3e4352a66f8faa5f324cb8 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T03:43:02Z |
| publishDate | 2022-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-f870682d0d3e4352a66f8faa5f324cb82023-11-23T11:27:57ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-05-012310575510.3390/ijms23105755Application of CRISPR/CasΦ2 System for Genome Editing in PlantsQinan Cai0Dongmei Guo1Yujun Cao2Yuan Li3Rui Ma4Wenping Liu5Maize Resources Institute, Jilin Academy of Agricultural Sciences, Gongzhuling 136100, ChinaInstitute of Agro-Product Safety and Nutrition, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, ChinaInstitute of Agricultural Resources and Environment, Jilin Academy of Agricultural Sciences, Changchun 130033, ChinaKey Laboratory of Agricultural Biotechnology, Jilin Academy of Agricultural Sciences, Changchun 130033, ChinaKey Laboratory of Agricultural Biotechnology, Jilin Academy of Agricultural Sciences, Changchun 130033, ChinaMaize Resources Institute, Jilin Academy of Agricultural Sciences, Gongzhuling 136100, ChinaCRISPR/Cas system has developed a new technology to modify target genes. In this study, CasΦ2 is a newly Cas protein that we used for genome modification in Arabidopsis and tobacco. <i>PDS</i> and <i>BRI1</i> of marker genes were chosen for targeting. CasΦ2 has the function to cleave pre-crRNA. In the presence of 10 mM Mg<sup>2+</sup> irons concentration, sgRNA3 type guided CasΦ2 to edit target gene and generate mutation, and a mutant seedling of <i>AtBRI1</i> gene with an expected male sterile phenotype was obtained. In the process of tobacco transformation, the gene editing activity of CasΦ2 can be activated by 100 nM Mg<sup>2+</sup> irons concentration, and sgRNA1 type guided CasΦ2 to edit target gene. Mutant seedlings of <i>NtPDS</i> gene with an expected albino were obtained. The results indicate that CasΦ2 can effectively edit target genes under the guidance of different sgRNA type in the presence of Mg<sup>2+</sup> ions. Together, our results verify that the CRISPR/CasΦ2 system is an effective and precise tool for genome editing in plants.https://www.mdpi.com/1422-0067/23/10/5755CRISPRCasΦ2genome editingMg<sup>2+</sup><i>Arabidopsis thaliana</i><i>Nicotiana benthamiana</i> |
| spellingShingle | Qinan Cai Dongmei Guo Yujun Cao Yuan Li Rui Ma Wenping Liu Application of CRISPR/CasΦ2 System for Genome Editing in Plants International Journal of Molecular Sciences CRISPR CasΦ2 genome editing Mg<sup>2+</sup> <i>Arabidopsis thaliana</i> <i>Nicotiana benthamiana</i> |
| title | Application of CRISPR/CasΦ2 System for Genome Editing in Plants |
| title_full | Application of CRISPR/CasΦ2 System for Genome Editing in Plants |
| title_fullStr | Application of CRISPR/CasΦ2 System for Genome Editing in Plants |
| title_full_unstemmed | Application of CRISPR/CasΦ2 System for Genome Editing in Plants |
| title_short | Application of CRISPR/CasΦ2 System for Genome Editing in Plants |
| title_sort | application of crispr casφ2 system for genome editing in plants |
| topic | CRISPR CasΦ2 genome editing Mg<sup>2+</sup> <i>Arabidopsis thaliana</i> <i>Nicotiana benthamiana</i> |
| url | https://www.mdpi.com/1422-0067/23/10/5755 |
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