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 and tested its capabilities in order to apply CRISPR/Cas9 technology for our purpose. In our experiments, we targeted two different genes, screened for mutation...

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Bibliographic Details
Published in:Frontiers in plant science 2018-06, Vol.9, p.740-740
Main Authors: Stukenberg, Daniel, Zauner, Stefan, Dell'Aquila, Gianluca, Maier, Uwe G
Format: Article
Language:English
Subjects:
CRISPR-Cas
diatom
genome editing
off-target
Phaeodactylum tricurnutum
Plant Science
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Summary:CRISPR/Cas9 is a powerful tool for genome editing. We constructed an easy-to-handle expression vector for application in the model organism 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 is very promising, although, as discussed, overlooked pitfalls have to be considered.
ISSN:1664-462X
1664-462X
DOI:10.3389/fpls.2018.00740