ArticleNature communications2018
One-step generation of multiple gene knock-outs in the diatom Phaeodactylum tricornutum by DNA-free genome editing.
Article in Nature communications, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 55 papers.
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Who cites it
55 citing papers in PubMed, 154 citations in OpenAlex.
- CRISPR-enhanced microalgae for carbon sequestration and carotenoid production: opportunities and fundamental constraints.Bioresources and bioprocessing · 2026Review
- CRISPR RNP-Mediated Transgene-Free Genome Editing in Plants: Advances, Challenges and Future Directions for Tree Species.Plant, cell & environment · 2026Review
- Enhancing recombinant eGFP secretion in P. tricornutum by optimizing culture conditions.Microbial cell factories · 2026Article
- NPF2 is involved in intracellular pH regulation and ion balance in the diatom Phaeodactylum tricornutum.The New phytologist · 2026Article
- Insights into transcriptomics and metabolic engineering of microalgal systems for enhancing industrial and environmental applications.World journal of microbiology & biotechnology · 2026Review
- PHYCUT: Scalable Multiplex CRISPR/Cas9 Editing for Genome Engineering in the DiatomACS synthetic biology · 2026Article
- Highly efficient genome editing using CRISPR/Cas9 ribonucleoprotein in the marine oleaginous diatom Fistulifera solaris.Scientific reports · 2026Article
- Review
- Breaking the cell wall for efficient DNA delivery to diatoms.Nature communications · 2026Article
- The marine diatom Phaeodactylum tricornutum as a versatile bioproduction chassis: Current progress, challenges, and perspectives.Plant communications · 2025Review
- A new tool for engineering Phaeodactylum tricornutum: the METE promoter drives both high expression and BThe Plant journal : for cell and molecular biology · 2025Article
- Tapping the microalgal potential: genetic precision and stress-induction for enhanced astaxanthin and biofuel production.Biotechnology for biofuels and bioproducts · 2025Review
- Circadian regulation of key physiological processes by the RITMO1 clock protein in the marine diatom Phaeodactylum tricornutum.The New phytologist · 2025Article
- Genetic engineering in diatoms: advances and prospects.The Plant journal : for cell and molecular biology · 2025Review
- Progress and challenges in CRISPR/Cas applications in microalgae.Journal of microbiology (Seoul, Korea) · 2025Review
- Marine Phytoplankton Bioactive Lipids and Their Perspectives in Clinical Inflammation.Marine drugs · 2025Review
- Article
- Biomanufacturing of γ-linolenic acid-enriched galactosyldiacylglycerols: Challenges in microalgae and potential in oleaginous yeasts.Synthetic and systems biotechnology · 2023Review
- Microbial applications for sustainable space exploration beyond low Earth orbit.NPJ microgravity · 2023Review
- CRISPR-based bioengineering in microalgae for production of industrially important biomolecules.Frontiers in bioengineering and biotechnology · 2023Review
Corrections and comments
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Authors and funding
6 authors at 4 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Recently developed transgenic techniques to explore and exploit the metabolic potential of microalgae present several drawbacks associated with the delivery of exogenous DNA into the cells and its subsequent integration at random sites within the genome. Here, we report a highly efficient multiplex genome-editing method in the diatom Phaeodactylum tricornutum, relying on the biolistic delivery of CRISPR-Cas9 ribonucleoproteins coupled with the identification of two endogenous counter-selectable markers, PtUMPS and PtAPT. First, we demonstrate the functionality of RNP delivery by positively selecting the disruption of each of these genes. Then, we illustrate the potential of the approach for multiplexing by generating double-gene knock-out strains, with 65% to 100% efficiency, using RNPs targeting one of these markers and PtAureo1a, a photoreceptor-encoding gene. Finally, we created triple knock-out strains in one step by delivering six RNP complexes into Phaeodactylum cells. This approach could readily be applied to other hard-to-transfect organisms of biotechnological interest.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.