Development of SyneBrick Vectors As a Synthetic Biology Platform for Gene Expression in Synechococcus elongatus PCC 7942
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Cyanobacteria are oxygenic photosynthetic prokaryotes that are able to assimilate CO2 using solar energy and water. Metabolic engineering of cyanobacteria has suggested the possibility of direct CO2 conversion to value-added chemicals. However, engineering of cyanobacteria has been limited due to the lack of various genetic tools for expression and control of multiple genes to reconstruct metabolic pathways for biochemicals from CO2. Thus, we developed SyneBrick vectors as a synthetic biology platform for gene expression in Synechococcus elongatus PCC 7942 as a model cyanobacterium. The SyneBrick chromosomal integration vectors provide three inducible expression systems to control gene expression and three neutral sites for chromosomal integrations. Using a SyneBrick vector, LacI-regulated gene expression led to 24-fold induction of the eYFP reporter gene with 1 mM isopropyl β-D-1-thiogalactopyranoside (IPTG) inducer in S. elongatus PCC 7942 under 5% (v/v) CO2. TetR-regulated gene expression led to 19-fold induction of the GFP gene when 100 nM anhydrotetracycline (aTc) inducer was used. Gene expression decreased after 48 h due to degradation of aTc under light. T7 RNA polymerase-based gene expression resulted in efficient expression with a lower IPTG concentration than a previously developed pTrc promoter. A library of T7 promoters can be used for tunable gene expression. In summary, SyneBrick vectors were developed as a synthetic biology platform for gene expression in S. elongatus PCC 7942. These results will accelerate metabolic engineering of biosolar cell factories through expressing and controlling multiple genes of interest.
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Neutral integration sites have been identified and characterised in commonly used cyanobacterial model organisms, such as Synechocystis sp. PCC 6803 and Synechococcus elongatus PCC 7942, providing reliable genomic locations for the stable insertion of heterologous genes and engineered metabolic pathways.
“Neutral integration sites have been identified and characterised in commonly used cyanobacterial model organisms, such as Synechocystis sp. PCC 6803 and Synechococcus elongatus PCC 7942, providing reliable genomic locations for the stable insertion of heterologous genes and engineered metabolic pathways [94,95].”
From Cyanobacteria as a Photosynthetic Chassis for Metabolic Pathway Engineering with Heterologous Gene Expression (2026) · cites this paper as [95]
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References 31 · Cited by 70
- Authors
- Kim, W. J., Lee, S.-M., Um, Y., Sim, S. J., & Woo, H. M.
- Affiliations
- Korea University (KR)
- Korea Institute of Science and Technology (KR)
- Sungkyunkwan University (KR)
- Corresponding author
- Woo, Han Min
- Venue
- Frontiers in Plant Science
- Publisher
- Frontiers Media SA
- Published
- 2017-03-02
- Volume
- 8
- Pages
- 293-293
- ISSN
- 1664-462X (ISSN-L)
- PMC
- PMC5332412
- OpenAlex
- W2595281090
- Cited by
- 70
- References
- 31
- Citations by year
- 2026: 2 · 2025: 5 · 2024: 4 · 2023: 5 · 2022: 8 · 2021: 9 · 2020: 16 · 2019: 10 · 2018: 7 · 2017: 4
- FWCI
- 4.13
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- cc-by
- Locations
- Frontiers in Plant Science (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Funders
- Ministry of Science, ICT and Future Planning (2014M1A8A1049277)
- National Research Foundation of Korea (2016, University-Institute Cooperation program)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Gene Regulatory Network Analysis
- Keywords
- Synechococcus elongatus PCC 7942, cyanobacteria, expression vectors, chromosomal integration, T7 RNA polymerase, IPTG, anhydrotetracycline, T7 promoter, metabolic engineering, CO2 conversion
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