Broad-host-range vector system for synthetic biology and biotechnology in cyanobacteria
Open accessAbstract
Inspired by the developments of synthetic biology and the need for improved genetic tools to exploit cyanobacteria for the production of renewable bioproducts, we developed a versatile platform for the construction of broad-host-range vector systems. This platform includes the following features: (i) an efficient assembly strategy in which modules released from 3 to 4 donor plasmids or produced by polymerase chain reaction are assembled by isothermal assembly guided by short GC-rich overlap sequences. (ii) A growing library of molecular devices categorized in three major groups: (a) replication and chromosomal integration; (b) antibiotic resistance; (c) functional modules. These modules can be assembled in different combinations to construct a variety of autonomously replicating plasmids and suicide plasmids for gene knockout and knockin. (iii) A web service, the CYANO-VECTOR assembly portal, which was built to organize the various modules, facilitate the in silico construction of plasmids, and encourage the use of this system. This work also resulted in the construction of an improved broad-host-range replicon derived from RSF1010, which replicates in several phylogenetically distinct strains including a new experimental model strain Synechocystis sp. WHSyn, and the characterization of nine antibiotic cassettes, four reporter genes, four promoters, and a ribozyme-based insulator in several diverse cyanobacterial strains.
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Suicide plasmids are widely used in targeted genome modification and gene knockout techniques within cyanobacterial genetic engineering.
“As such, suicide plasmids are widely used in targeted genome modification and gene knockout techniques within cyanobacterial genetic engineering [91,92].”
From Cyanobacteria as a Photosynthetic Chassis for Metabolic Pathway Engineering with Heterologous Gene Expression (2026) · cites this paper as [91]
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Xie, H., Bourgade, B., Stensjö, K., & Lindblad, P. (2025) · Microbial Cell Factories · doi:10.1186/s12934-025-02727-8 · Cited by 11 · see citing works
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“As such, suicide plasmids are widely used in targeted genome modification and gene knockout techniques within cyanobacterial genetic engineering [91,92].”
- Authors
- Taton, A., Unglaub, F., Wright, N. E., Zeng, W. Y., Paz-Yepes, J., Brahamsha, B., Palenik, B., Peterson, T. C., Haerizadeh, F., Golden, S. S., & Golden, J. W.
- Affiliations
- University of California San Diego (US)
- Centre National de la Recherche Scientifique (FR)
- Scripps Institution of Oceanography (US)
- Corresponding author
- Taton, Arnaud
- Venue
- Nucleic Acids Research
- Publisher
- Oxford University Press (OUP)
- Published
- 2014-07-29
- Volume
- 42
- Issue
- 17
- Pages
- e136-e136
- ISSN
- 0305-1048 (ISSN-L)
- 1362-4962
- PMC
- PMC4176158
- OpenAlex
- W2128556638
- Cited by
- 186
- References
- 76
- Citations by year
- 2026: 15 · 2025: 17 · 2024: 15 · 2023: 12 · 2022: 20 · 2021: 18 · 2020: 14 · 2019: 23 · 2018: 25 · 2017: 9 · 2016: 8 · 2015: 7 · 2014: 3
- FWCI
- 3.69
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- Nucleic Acids Research (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- eScholarship (California Digital Library) (submittedVersion)
- Funders
- Life Technologies Corporation
- Virginia Commonwealth University
- European Commission
- National Institute of General Medical Sciences (R01GM062419)
- 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 › CRISPR and Genetic Engineering
- Keywords
- broad-host-range vectors, cyanobacteria, plasmid assembly, suicide plasmid, antibiotic resistance cassettes, reporter genes, promoter, plasmid replication, chromosomal integration, molecular devices
- MeSH terms
- Cyanobacteria, Biotechnology, Computer Simulation, Drug Resistance, Microbial, Genetic Vectors, Plasmids, Replicon, Genes, Reporter, Gene Targeting, Synthetic Biology
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