AUTHOR=Stukenberg Daniel , Zauner Stefan , Dell’Aquila Gianluca , Maier Uwe G. TITLE=Optimizing CRISPR/Cas9 for the Diatom Phaeodactylum tricornutum JOURNAL=Frontiers in Plant Science VOLUME=9 YEAR=2018 URL=https://www.frontiersin.org/journals/plant-science/articles/10.3389/fpls.2018.00740 DOI=10.3389/fpls.2018.00740 ISSN=1664-462X 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.