ArticleScientific reports2022
Phosphate-regulated expression of the SARS-CoV-2 receptor-binding domain in the diatom Phaeodactylum tricornutum for pandemic diagnostics.
Article in Scientific reports, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 18 papers, 1 of them a synthesis that pooled it.
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Who cites it
18 citing papers in PubMed, 1 synthesis or guideline pooled it, 33 citations in OpenAlex.
- Meta-analysis of RNA-Seq datasets allows a better understanding of P. tricornutum cellular biology, a requirement to improve the production of Biologics.Scientific reports · 2025Pooled it
- Enhancing recombinant eGFP secretion in P. tricornutum by optimizing culture conditions.Microbial cell factories · 2026Article
- PHYCUT: Scalable Multiplex CRISPR/Cas9 Editing for Genome Engineering in the DiatomACS synthetic biology · 2026Article
- Breaking the cell wall for efficient DNA delivery to diatoms.Nature communications · 2026Article
- Development of a semi-endogenous PRT-based expression system for Methylotuvimicrobium alcaliphilum and its application to enhance 3-hydroxybutyric acid production from methanol.Journal of biological engineering · 2025Review
- The marine diatom Phaeodactylum tricornutum as a versatile bioproduction chassis: Current progress, challenges, and perspectives.Plant communications · 2025Review
- A game of tag: A review of protein tags for the successful detection, purification and fluorescence labelling of proteins expressed in microalgae.The Plant journal : for cell and molecular biology · 2025Review
- Characterization of the VOC Promoter That Is Active Under Low-Salinity Conditions in the DiatomMarine drugs · 2025Article
- In Vitro Culture, Genetic Transformation and the Production of Biopharmaceuticals in Microalgae.International journal of molecular sciences · 2025Review
- Genetic engineering in diatoms: advances and prospects.The Plant journal : for cell and molecular biology · 2025Review
- Metabolic response to a heterologous poly-3-hydroxybutyrate (PHB) pathway in Phaeodactylum tricornutum.Applied microbiology and biotechnology · 2024Article
- Review
- No two clones are alike: characterization of heterologous subpopulations in a transgenic cell line of the model diatom Phaeodactylum tricornutum.Microbial cell factories · 2024Article
- In vivo thrombin activity in the diatom Phaeodactylum tricornutum: biotechnological insights.Applied microbiology and biotechnology · 2024Article
- Comparative RNA-Seq of TenMarine drugs · 2024Article
- Review
- Microalgae as an Efficient Vehicle for the Production and Targeted Delivery of Therapeutic Glycoproteins against SARS-CoV-2 Variants.Marine drugs · 2022Review
- Production and secretion of functional SARS-CoV-2 spike protein inFrontiers in plant science · 2022Article
Corrections and comments
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Authors and funding
24 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The worldwide COVID-19 pandemic caused by the SARS-CoV-2 betacoronavirus has highlighted the need for a synthetic biology approach to create reliable and scalable sources of viral antigen for uses in diagnostics, therapeutics and basic biomedical research. Here, we adapt plasmid-based systems in the eukaryotic microalgae Phaeodactylum tricornutum to develop an inducible overexpression system for SARS-CoV-2 proteins. Limiting phosphate and iron in growth media induced expression of the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein from the P. tricornutum HASP1 promoter in the wild-type strain and in a histidine auxotrophic strain that alleviates the requirement for antibiotic selection of expression plasmids. The RBD was purified from whole cell extracts (algae-RBD) with yield compromised by the finding that 90-95% of expressed RBD lacked the genetically encoded C-terminal 6X-histidine tag. Constructs that lacked the TEV protease site between the RBD and C-terminal 6X-histidine tag retained the tag, increasing yield. Purified algae-RBD was found to be N-linked glycosylated by treatment with endoglycosidases, was cross-reactive with anti-RBD polyclonal antibodies, and inhibited binding of recombinant RBD purified from mammalian cell lines to the human ACE2 receptor. We also show that the algae-RBD can be used in a lateral flow assay device to detect SARS-CoV-2 specific IgG antibodies from donor serum at sensitivity equivalent to assays performed with RBD made in mammalian cell lines. Our study shows that P. tricornutum is a scalable system with minimal biocontainment requirements for the inducible production of SARS-CoV-2 or other coronavirus antigens for pandemic diagnostics.
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