ArticlePlant biotechnology journal2016
Codon reassignment to facilitate genetic engineering and biocontainment in the chloroplast of Chlamydomonas reinhardtii.
Article in Plant biotechnology journal, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
What it found
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
22 citing papers in PubMed, 51 citations in OpenAlex.
- Article
- Article
- Microalgae for bioremediation: advances, challenges, and public perception on genetic engineering.BMC plant biology · 2024Review
- The synthetic future of algal genomes.Cell genomics · 2024Review
- Harnessing the Algal Chloroplast for Heterologous Protein Production.Microorganisms · 2022Review
- Overview of tRNA Modifications in Chloroplasts.Microorganisms · 2022Article
- Biocontainment of Genetically Engineered Algae.Frontiers in plant science · 2022Review
- Microbial Biocontainment Systems for Clinical, Agricultural, and Industrial Applications.Frontiers in bioengineering and biotechnology · 2022Review
- The Algal Chloroplast as a Testbed for Synthetic Biology Designs Aimed at Radically Rewiring Plant Metabolism.Frontiers in plant science · 2021Review
- A microalgal-based preparation with synergistic cellulolytic and detoxifying action towards chemical-treated lignocellulose.Plant biotechnology journal · 2021Article
- Nanotechnology Approaches for Chloroplast Biotechnology Advancements.Frontiers in plant science · 2021Review
- Multigenic engineering of the chloroplast genome in the green algaMicrobiology (Reading, England) · 2020Article
- The phosphite oxidoreductase gene, ptxD as a bio-contained chloroplast marker and crop-protection tool for algal biotechnology using Chlamydomonas.Applied microbiology and biotechnology · 2020Article
- Assessment of biomass potentials of microalgal communities in open pond raceways using mass cultivation.PeerJ · 2020Article
- Use of the ptxD gene as a portable selectable marker for chloroplast transformation in Chlamydomonas reinhardtii.Molecular biotechnology · 2019Article
- CITRIC: cold-inducible translational readthrough in the chloroplast of Chlamydomonas reinhardtii using a novel temperature-sensitive transfer RNA.Microbial cell factories · 2018Article
- Review
- Review
- Green biologics: The algal chloroplast as a platform for making biopharmaceuticals.Bioengineered · 2018Review
- Synthetic biology approaches to biological containment: pre-emptively tackling potential risks.Essays in biochemistry · 2016Review
Corrections and comments
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Authors and funding
2 authors at 2 institutions in 1 country.
Funding
Abstract
There is a growing interest in the use of microalgae as low-cost hosts for the synthesis of recombinant products such as therapeutic proteins and bioactive metabolites. In particular, the chloroplast, with its small, genetically tractable genome (plastome) and elaborate metabolism, represents an attractive platform for genetic engineering. In Chlamydomonas reinhardtii, none of the 69 protein-coding genes in the plastome uses the stop codon UGA, therefore this spare codon can be exploited as a useful synthetic biology tool. Here, we report the assignment of the codon to one for tryptophan and show that this can be used as an effective strategy for addressing a key problem in chloroplast engineering: namely, the assembly of expression cassettes in Escherichia coli when the gene product is toxic to the bacterium. This problem arises because the prokaryotic nature of chloroplast promoters and ribosome-binding sites used in such cassettes often results in transgene expression in E. coli, and is a potential issue when cloning genes for metabolic enzymes, antibacterial proteins and integral membrane proteins. We show that replacement of tryptophan codons with the spare codon (UGG→UGA) within a transgene prevents functional expression in E. coli and in the chloroplast, and that co-introduction of a plastidial trnW gene carrying a modified anticodon restores function only in the latter by allowing UGA readthrough. We demonstrate the utility of this system by expressing two genes known to be highly toxic to E. coli and discuss its value in providing an enhanced level of biocontainment for transplastomic microalgae.
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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.