Original Research ARTICLE
Optimizing CRISPR/Cas9 for the diatom Phaeodactylum tricornutum
- 1Cell Biology, Philipps University of Marburg, Germany
- 2LOEWE Zentrum für synthetische Mikrobiologie (SYNMIKRO), Philipps University of Marburg, Germany
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.
Keywords: CRISPR-Cas, Genome editing, Phaeodactylum tricurnutum, diatom, off-target
Received: 15 Jan 2018;
Accepted: 15 May 2018.
Edited by:Roger Deal, Emory University, United States
Reviewed by:Per Winge, Norwegian University of Science and Technology, Norway
Fayza Daboussi, Institut National de la Recherche Agronomique (INRA), France
Copyright: © 2018 Stukenberg, Zauner, Dell'Aquila and Maier. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Prof. Uwe G. Maier, UGM., Philipps University of Marburg, Cell Biology, Marburg, Germany, firstname.lastname@example.org