ArticleApplied microbiology and biotechnology2016
New tools for chloroplast genetic engineering allow the synthesis of human growth hormone in the green alga Chlamydomonas reinhardtii.
Article in Applied microbiology and biotechnology, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 51 papers.
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51 citing papers in PubMed, 115 citations in OpenAlex.
- Recombinant bovine chymosin expression in microalgae Chlamydomonas reinhardtii chloroplast: A step towards algal biomanufacturing of dairy enzymes.World journal of microbiology & biotechnology · 2026Article
- Process intensification for the protein production from microalgae: a comprehensive review.Bioresources and bioprocessing · 2026Review
- Partitioning of Rubisco activase into the pyrenoidal Rubisco condensate is mediated by a functional protein-protein interaction.Nature communications · 2026Article
- Engineering bidirectional chloroplast promoters for tunable co-expression of multiple genes in microalgae (Chlamydomonas reinhardtii).Communications biology · 2026Article
- A modular high-throughput approach for advancing synthetic biology in the chloroplast of Chlamydomonas.Nature plants · 2025Article
- The First Introduction of an Exogenous 5' Untranslated Region for Control of Plastid Transgene Expression in Chlamydomonas reinhardtii.Molecular biotechnology · 2025Article
- Genetically engineered bacteria and microalgae expressing a mutant of cytochrome P450 BM3 for efficient Diuron degradation in wastewater treatment.Microbiology spectrum · 2025Article
- 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
- Exploring microalgae-based strategies for the management of white spot syndrome virus: a comprehensive review.3 Biotech · 2025Review
- In Vitro Culture, Genetic Transformation and the Production of Biopharmaceuticals in Microalgae.International journal of molecular sciences · 2025Review
- Epidemiological analysis of turner syndrome in children aged 0-14 years: global, regional, and national perspectives (1990-2021).Frontiers in endocrinology · 2025Article
- Heterologous Gene Expression in Chlamydomonas reinhardtii Chloroplast by Heterologous Promoters and Terminators, Intercistronic Expression Elements and Minichromosome.Microbial biotechnology · 2024Article
- Harnessing the potential of microalgae for the production of monoclonal antibodies and other recombinant proteins.Protoplasma · 2024Review
- Pharmaceutical removal from wastewater by introducing cytochrome P450s into microalgae.Microbial biotechnology · 2024Review
- Review
- The synthetic future of algal genomes.Cell genomics · 2024Review
- An RNA thermometer in the chloroplast genome of Chlamydomonas facilitates temperature-controlled gene expression.Nucleic acids research · 2023Article
- Accelerating Chloroplast Engineering: A New System for Rapid Generation of Marker-Free Transplastomic Lines ofMicroorganisms · 2023Article
- Transgenic Microalgae Expressing Double-Stranded RNA as Potential Feed Supplements for Controlling White Spot Syndrome in Shrimp Aquaculture.Microorganisms · 2023Article
- A PETase enzyme synthesised in the chloroplast of the microalga Chlamydomonas reinhardtii is active against post-consumer plastics.Scientific reports · 2023Article
Corrections and comments
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Authors and funding
5 authors at 3 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
In recent years, there has been an increasing interest in the exploitation of microalgae in industrial biotechnology. Potentially, these phototrophic eukaryotes could be used for the low-cost synthesis of valuable recombinant products such as bioactive metabolites and therapeutic proteins. The algal chloroplast in particular represents an attractive target for such genetic engineering, both because it houses major metabolic pathways and because foreign genes can be targeted to specific loci within the chloroplast genome, resulting in high-level, stable expression. However, routine methods for chloroplast genetic engineering are currently available only for one species-Chlamydomonas reinhardtii-and even here, there are limitations to the existing technology, including the need for an expensive biolistic device for DNA delivery, the lack of robust expression vectors, and the undesirable use of antibiotic resistance markers. Here, we describe a new strain and vectors for targeted insertion of transgenes into a neutral chloroplast locus that (i) allow scar-less fusion of a transgenic coding sequence to the promoter/5'UTR element of the highly expressed endogenous genes psaA or atpA, (ii) employ the endogenous gene psbH as an effective but benign selectable marker, and (iii) ensure the successful integration of the transgene construct in all transformant lines. Transformation is achieved by a simple and cheap method of agitation of a DNA/cell suspension with glass beads, with selection based on the phototrophic rescue of a cell wall-deficient ΔpsbH strain. We demonstrate the utility of these tools in the creation of a transgenic line that produces high levels of functional human growth hormone.
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