Abstract
Diatoms are major components of phytoplankton and play a key role in the ecology of aquatic ecosystems. These algae are of great scientific importance for a wide variety of research areas, ranging from marine ecology and oceanography to biotechnology. During the last 20 years, the availability of genomic information on selected diatom species and a substantial progress in genetic manipulation, strongly contributed to establishing diatoms as molecular model organisms for marine biology research. Recently, tailored TALEN endonucleases and the CRISPR/Cas9 system were utilized in diatoms, allowing targeted genetic modifications and the generation of knockout strains. These approaches are extremely valuable for diatom research because breeding, forward genetic screens by random insertion, and chemical mutagenesis are not applicable to the available model species Phaeodactylum tricornutum and Thalassiosira pseudonana, which do not cross sexually in the lab. Here, we provide an overview of the genetic toolbox that is currently available for performing stable genetic modifications in diatoms. We also discuss novel challenges that need to be addressed to fully exploit the potential of these technologies for the characterization of diatom biology and for metabolic engineering.
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Acknowledgements
The authors would like to thank the Gordon and Betty Moore Foundation for generously supporting the project GBMF 4966 “DiaEdit—Development of genetic tools for the establishment of routine genome editing in the marine diatom Phaeodactylum tricornutum”. Research on genome editing in AF and MF labs is also supported by the European Assemble plus (Association of European Marine Biological Research Laboratories Expanded, H2020-INFRAIA-1-2016-2017) consortium. Several vectors for P. tricornutum genome editing generated in the framework of the DiaEdit consortium are available through Addgene: the CRISPR-Cas9 pKS_diaCas9_sgRNA (Addgene ID: 74923, https://www.addgene.org/74923/), the pPtPuc3m_diaCas9_sgRNA for episome-based genome editing (Addgene ID: 109219, https://www.addgene.org/109219/) (Winge et al. unpublished), and 8 TALEN Vectors (Addgene ID: 90415–90423; https://www.addgene.org/Peter_Kroth/).
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Communicated by Neal Stewart.
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Kroth, P.G., Bones, A.M., Daboussi, F. et al. Genome editing in diatoms: achievements and goals. Plant Cell Rep 37, 1401–1408 (2018). https://doi.org/10.1007/s00299-018-2334-1
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DOI: https://doi.org/10.1007/s00299-018-2334-1