ArticleNucleic acids research2018
Intron-containing algal transgenes mediate efficient recombinant gene expression in the green microalga Chlamydomonas reinhardtii.
Article in Nucleic acids research, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 71 papers.
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71 citing papers in PubMed.
- Transfer of chloroplastSynthetic and systems biotechnology · 2027Article
- Heme-binding protein CYB5D1 couples intraflagellar redox to calcium signaling for coordinated flagellar beating.Proceedings of the National Academy of Sciences of the United States of America · 2026Article
- Over-Expression of a Phycocyanin-Interferon Fusion and Differential Cleaving Efficiency in Cyanobacteria (Synechocystis sp. PCC 6803).Biotechnology and bioengineering · 2026Article
- Synthetic Biology and Metabolic Engineering of Microalgae for Sustainable Lipid and Terpenoid Production: An Updated Perspective.Plant biotechnology journal · 2026Review
- Pre-mRNA splicing and its regulation in microalgae and cyanobacteria.Advanced biotechnology · 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
- Sustainable Production of Bio-Based Geraniol: Heterologous Expression of Early Terpenoid Pathway Enzymes inACS synthetic biology · 2025Article
- Transcriptional gene fusions via targeted integration at safe harbors for high transgene expression in Chlamydomonas reinhardtii.The New phytologist · 2025Article
- Programmed cell death and redox metabolism protect Chlamydomonas reinhardtii populations from the galactic cosmic environment on the Artemis-1 mission.Scientific reports · 2025Article
- Complexome profiling of the Chlamydomonas psb28 mutant reveals TEF5 as an early PSII assembly factor.The Plant cell · 2025Article
- Effect of recombinant protein production and release on microalgal fitness and the impact of environmental conditions for localized therapeutic delivery.Journal of biological engineering · 2025Article
- EngineeredLife (Basel, Switzerland) · 2025Article
- Genome editing in the green alga Chlamydomonas: past, present practice and future prospects.The Plant journal : for cell and molecular biology · 2025Review
- Internalization of affinity tags enables the purification of secreted Chlamydomonas proteins.Current genetics · 2025Article
- Solving Challenges in Microalgae-Based Living Materials.ACS synthetic biology · 2025Article
- Mechanism of Transcription Factor ChbZIP1 Enhanced Alkaline Stress Tolerance inInternational journal of molecular sciences · 2025Article
- A novel MoClo-mediated intron insertion system facilitates enhanced transgene expression inFrontiers in plant science · 2025Article
- Golden Gate Cloning for the Standardized Assembly of Gene Elements with Modular Cloning in Chlamydomonas.Methods in molecular biology (Clifton, N.J.) · 2025Article
- Fine-tuning the N-glycosylation of recombinant human erythropoietin using Chlamydomonas reinhardtii mutants.Plant biotechnology journal · 2024Article
- Proteomic analysis reveals molecular changes following genetic engineering in Chlamydomonas reinhardtii.BMC microbiology · 2024Article
11 more citing papers are in PubMed but not listed here.
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4 authors.
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Abstract
Among green freshwater microalgae, Chlamydomonas reinhardtii has the most comprehensive and developed molecular toolkit, however, advanced genetic and metabolic engineering driven from the nuclear genome is generally hindered by inherently low transgene expression levels. Progressive strain development and synthetic promoters have improved the capacity of transgene expression; however, the responsible regulatory mechanisms are still not fully understood. Here, we elucidate the sequence specific dynamics of native regulatory element insertion into nuclear transgenes. Systematic insertions of the first intron of the ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit 2 (rbcS2i1) throughout codon-optimized coding sequences (CDS) generates optimized algal transgenes which express reliably in C. reinhardtii. The optimal rbcS2i1 insertion site for efficient splicing was systematically determined and improved gene expression rates were shown using a codon-optimized sesquiterpene synthase CDS. Sequential insertions of rbcS2i1 were found to have a step-wise additive effect on all levels of transgene expression, which is likely correlated to a synergy of transcriptional machinery recruitment and mimicking the short average exon lengths natively found in the C. reinhardtii genome. We further demonstrate the value of this optimization with five representative transgene examples and provide guidelines for the design of any desired sequence with this strategy.
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