Targeted CRISPR‐Cas9‐based gene knockouts in the model brown alga Ectocarpus
Badis, Y.; Scornet, D.; Harada, M.; Caillard, C.; Godfroy, O.; Raphalen, M.; Gachon, C.M.M.; Coelho, S.M.; Motomura, T.; Nagasato, C.; Cock, J.M. (2021). Targeted CRISPR‐Cas9‐based gene knockouts in the model brown alga Ectocarpus. New Phytol. 231(5): 2077-2091. https://dx.doi.org/10.1111/nph.17525
In: New Phytologist. Wiley-Blackwell: Oxford. ISSN 0028-646X; e-ISSN 1469-8137, meer
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| Trefwoorden |
ASSEMBLEPlus Joint Research Activity 3
Exploitable Scientific Result
Scientific Publication
Ectocarpus Lyngbye, 1819 [WoRMS]
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| Author keywords |
brown alga, Cas9, CRISPR, everse genetics, transformation |
| Auteurs | | Top |
- Badis, Y.
- Scornet, D.
- Harada, M.
- Caillard, C.
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- Godfroy, O.
- Raphalen, M.
- Gachon, C.M.M.
- Coelho, S.M.
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- Motomura, T.
- Nagasato, C.
- Cock, J.M.
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| Abstract |
Brown algae are an important group of multicellular eukaryotes, phylogenetically distinct from both the animal and land plant lineages. Ectocarpus has emerged as a model organism to study diverse aspects of brown algal biology, but this system currently lacks an effective reverse genetics methodology to analyse the functions of selected target genes. - Here, we report that mutations at specific target sites are generated following the introduction of CRISPR-Cas9 ribonucleoproteins into Ectocarpus cells, using either biolistics or microinjection as the delivery method.
- Individuals with mutations affecting the ADENINE PHOSPHORIBOSYL TRANSFERASE (APT) gene were isolated following treatment with 2-fluoroadenine, and this selection system was used to isolate individuals in which mutations had been introduced simultaneously at APT and at a second gene. This double mutation approach could potentially be used to isolate mutants affecting any Ectocarpus gene, providing an effective reverse genetics tool for this model organism.
- The availability of this tool will significantly enhance the utility of Ectocarpus as a model organism for this ecologically and economically important group of marine organisms. Moreover, the methodology described here should be readily transferable to other brown algal species.
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