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Optimizing CRISPR/Cas9 for the Diatom Phaeodactylum tricornutum.

ABSTRACT: 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.

SUBMITTER: Stukenberg D

PROVIDER: S-EPMC5998643 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature

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Optimizing CRISPR/Cas9 for the Diatom <i>Phaeodactylum tricornutum</i>.

Stukenberg Daniel D Zauner Stefan S Dell'Aquila Gianluca G Maier Uwe G UG

Frontiers in plant science 20180606

CRISPR/Cas9 is a powerful tool for genome editing. We constructed an easy-to-handle expression vector for application in the model organism <i>Phaeodactylum tricornutum</i> 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 conf ...[more]

PMID: 29928285

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