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 WILD MANUKA OIL 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 Windows pitfalls have to be considered.