ArticleScientific reports2018
Transgene-free genome editing in marine algae by bacterial conjugation - comparison with biolistic CRISPR/Cas9 transformation.
Article in Scientific reports, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 31 papers.
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Who cites it
31 citing papers in PubMed.
- Roadmap for light interaction with biophotonic surfaces and their diverse applications.Journal of biomedical optics · 2026Article
- Microalgal Synthetic Biotechnology: From Genetic Models to Deployable Photosynthetic Chassis.Research (Washington, D.C.) · 2026Review
- Creating an Improved Diatoxanthin Production Line by Knocking OutMarine drugs · 2025Article
- Both major xanthophyll cycles present in nature promote nonphotochemical quenching in a model diatom.Plant physiology · 2025Article
- Genetic engineering in diatoms: advances and prospects.The Plant journal : for cell and molecular biology · 2025Review
- Tuning interdomain conjugation to enablemSystems · 2024Article
- Revolutionizing Lung Cancer Treatment: Innovative CRISPR-Cas9 Delivery Strategies.AAPS PharmSciTech · 2024Review
- Article
- A synthetic biology approach for the treatment of pollutants with microalgae.Frontiers in bioengineering and biotechnology · 2024Review
- Validating a Promoter Library for Application in Plasmid-Based Diatom Genetic Engineering.ACS synthetic biology · 2023Article
- Loss of CpFTSY Reduces Photosynthetic Performance and Affects Insertion of PsaC of PSI in Diatoms.Plant & cell physiology · 2023Article
- Microalgal Carotenoids: Therapeutic Application and Latest Approaches to Enhance the Production.Current issues in molecular biology · 2022Review
- Green diatom mutants reveal an intricate biosynthetic pathway of fucoxanthin.Proceedings of the National Academy of Sciences of the United States of America · 2022Article
- Phosphate-regulated expression of the SARS-CoV-2 receptor-binding domain in the diatom Phaeodactylum tricornutum for pandemic diagnostics.Scientific reports · 2022Article
- Genetic Engineering of Microalgae for Secondary Metabolite Production: Recent Developments, Challenges, and Future Prospects.Frontiers in bioengineering and biotechnology · 2022Review
- Impact of Lhcx2 on Acclimation to Low Iron Conditions in the DiatomFrontiers in plant science · 2022Article
- Telomere-to-telomere genome assembly ofPeerJ · 2022Article
- Method Development Progress in Genetic Engineering of Thraustochytrids.Marine drugs · 2021Review
- Simultaneous knockout of multiple LHCF genes using single sgRNAs and engineering of a high-fidelity Cas9 for precise genome editing in marine algae.Plant biotechnology journal · 2021Article
- SPX-related genes regulate phosphorus homeostasis in the marine phytoplankton, Phaeodactylum tricornutum.Communications biology · 2021Article
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Authors and funding
5 authors.
Funding
Abstract
The CRISPR/Cas9 technology has opened the possibility for targeted genome editing in various organisms including diatom model organisms. One standard method for delivery of vectors to diatom cells is by biolistic particle bombardment. Recently delivery by conjugation was added to the tool-box. An important difference between these methods is that biolistic transformation results in transgene integration of vector DNA into the algae genome, whereas conjugative transformation allows the vector to be maintained as an episome in the recipient cells. In this study, we have used both transformation methods to deliver the CRISPR/Cas9 system to the marine diatom Phaeodactylum tricornutum aiming to induce mutations in a common target gene. This allowed us to compare the two CRISPR/Cas9 delivery systems with regard to mutation efficiency, and to assess potential problems connected to constitutive expression of Cas9. We found that the percentage of CRISPR-induced targeted biallelic mutations are similar for both methods, but an extended growth period might be needed to induce biallelic mutations when the CRISPR/Cas9 system is episomal. Independent of the CRISPR/Cas9 vector system, constitutive expression of Cas9 can cause re-editing of mutant lines with small indels. Complications associated with the biolistic transformation system like the permanent and random integration of foreign DNA into the host genome and unstable mutant lines caused by constitutive expression of Cas9 can be avoided using the episomal CRISPR/Cas9 system. The episomal vector can be eliminated from the diatom cells by removal of selection pressure, resulting in transient Cas9 expression and non-transgenic mutant lines. Depending on legislation, such lines might be considered as non-GMOs.
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