Optimizing CRISPR/Cas9 for the Diatom Phaeodactylum tricornutum
CRISPR/Cas9 is a powerful tool for genome editing. We constructed an easy-to-handle expression vector for application in the model organism Phaeodactylum tricornutum and tested its capabilities in order to apply CRISPR/Cas9 technology for our purpose. In our experiments, we targeted two different ge...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2018-06-01
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| Series: | Frontiers in Plant Science |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/article/10.3389/fpls.2018.00740/full |
| _version_ | 1818199607301111808 |
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| author | Daniel Stukenberg Stefan Zauner Gianluca Dell’Aquila Uwe G. Maier Uwe G. Maier |
| author_facet | Daniel Stukenberg Stefan Zauner Gianluca Dell’Aquila Uwe G. Maier Uwe G. Maier |
| author_sort | Daniel Stukenberg |
| collection | DOAJ |
| description | CRISPR/Cas9 is a powerful tool for genome editing. We constructed an easy-to-handle expression vector for application in the model organism Phaeodactylum tricornutum and tested its capabilities in order to apply CRISPR/Cas9 technology for our purpose. In our experiments, we targeted two different genes, screened for mutations and analyzed mutated diatoms in a three-step process. In the end, we identified cells, showing either monoallelic or homo-biallelic targeted mutations. Thus, we confirm that application of the CRISPR/Cas9 system for P. tricornutum is very promising, although, as discussed, overlooked pitfalls have to be considered. |
| first_indexed | 2024-12-12T02:24:27Z |
| format | Article |
| id | doaj.art-06e830e686fd4cb7a24f6ffabddfcd65 |
| institution | Directory Open Access Journal |
| issn | 1664-462X |
| language | English |
| last_indexed | 2024-12-12T02:24:27Z |
| publishDate | 2018-06-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Plant Science |
| spelling | doaj.art-06e830e686fd4cb7a24f6ffabddfcd652022-12-22T00:41:36ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2018-06-01910.3389/fpls.2018.00740350635Optimizing CRISPR/Cas9 for the Diatom Phaeodactylum tricornutumDaniel Stukenberg0Stefan Zauner1Gianluca Dell’Aquila2Uwe G. Maier3Uwe G. Maier4Department for Cell Biology, Philipps-Universität Marburg, Marburg, GermanyDepartment for Cell Biology, Philipps-Universität Marburg, Marburg, GermanyDepartment for Cell Biology, Philipps-Universität Marburg, Marburg, GermanyDepartment for Cell Biology, Philipps-Universität Marburg, Marburg, GermanyLOEWE-Zentrum für Synthetische Mikrobiologie (SYNMIKRO), Philipps-Universität Marburg, Marburg, GermanyCRISPR/Cas9 is a powerful tool for genome editing. We constructed an easy-to-handle expression vector for application in the model organism Phaeodactylum tricornutum and tested its capabilities in order to apply CRISPR/Cas9 technology for our purpose. In our experiments, we targeted two different genes, screened for mutations and analyzed mutated diatoms in a three-step process. In the end, we identified cells, showing either monoallelic or homo-biallelic targeted mutations. Thus, we confirm that application of the CRISPR/Cas9 system for P. tricornutum is very promising, although, as discussed, overlooked pitfalls have to be considered.https://www.frontiersin.org/article/10.3389/fpls.2018.00740/fullCRISPR-Casgenome editingPhaeodactylum tricurnutumdiatomoff-target |
| spellingShingle | Daniel Stukenberg Stefan Zauner Gianluca Dell’Aquila Uwe G. Maier Uwe G. Maier Optimizing CRISPR/Cas9 for the Diatom Phaeodactylum tricornutum Frontiers in Plant Science CRISPR-Cas genome editing Phaeodactylum tricurnutum diatom off-target |
| title | Optimizing CRISPR/Cas9 for the Diatom Phaeodactylum tricornutum |
| title_full | Optimizing CRISPR/Cas9 for the Diatom Phaeodactylum tricornutum |
| title_fullStr | Optimizing CRISPR/Cas9 for the Diatom Phaeodactylum tricornutum |
| title_full_unstemmed | Optimizing CRISPR/Cas9 for the Diatom Phaeodactylum tricornutum |
| title_short | Optimizing CRISPR/Cas9 for the Diatom Phaeodactylum tricornutum |
| title_sort | optimizing crispr cas9 for the diatom phaeodactylum tricornutum |
| topic | CRISPR-Cas genome editing Phaeodactylum tricurnutum diatom off-target |
| url | https://www.frontiersin.org/article/10.3389/fpls.2018.00740/full |
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