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Improving the Transformation Efficiency of Synechococcus sp. PCC 7002 via Methylome-Guided Premethylation of DNA
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High Resolution Image Download MS PowerPoint Slide Cyanobacteria are promising microbial platforms for a diverse set of biotechnology applications, from living materials to photosynthetic chemical production, but are less well characterized than commonly engineered microbes such as Escherichia coli . This study facilitates genetic engineering in Synechococcus sp. PCC 7002, a fast-growing, halotolerant, and naturally competent strain, by identifying ten native methylation motifs and designing shuttle strains that mimic the native methylation state by expressing a subset of heterologous methyltransferases. DNA methylation in E. coli with as few as two active methyltransferases increased transformation efficiency up to 30-fold across four distinct integration sites in PCC 7002. This work provides an experimental framework to bypass native restriction-modification systems for efficient genome editing and metabolic engineering in nonmodel bacteria.
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Despite these obstacles, electroporation remains an important technique for introducing plasmid DNA into specific cyanobacterial strains, where alternative methods may be less effective.
“Despite these obstacles, electroporation remains an important technique for introducing plasmid DNA into specific cyanobacterial strains, where alternative methods may be less effective [85,86].”
From Cyanobacteria as a Photosynthetic Chassis for Metabolic Pathway Engineering with Heterologous Gene Expression (2026) · cites this paper as [85]
The Methods describe electroporation of modified BW25113 cloning strains, not cyanobacteria. The Introduction and the Methods section on PCC 7002 instead describe natural transformation with plasmid DNA. Our copy does not address whether electroporation is important for particular cyanobacterial strains or whether alternative methods are less effective for them. (LLM, conf=0.77)
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Metadata
- Authors
- Hren, A., Alexander, W. G., Abraham, J. P., Tumen-Velasquez, M. P., Melesse Vergara, M., Guss, A. M., Pfleger, B. F., Fox, J. M., & Eckert, C. A.
- Affiliations
- University of Colorado Boulder (US)
- Oak Ridge National Laboratory (US)
- University of Wisconsin System (US)
- Biosciences Division
- Corresponding author
- Pfleger, Brian F.
- Fox, Jerome M.
- Eckert, Carrie A.
- ORCID
- Venue
- ACS Synthetic Biology
- Publisher
- American Chemical Society (ACS)
- Published
- 2025-08-04
- Volume
- 14
- Issue
- 8
- Pages
- 3258-3264
- ISSN
- 2161-5063 (ISSN-L)
- PMC
- PMC12362590
- OpenAlex
- W4412895138
- Cited by
- 5
- References
- 40
- Citations by year
- 2026: 4 · 2025: 1
- FWCI
- 1.49
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- ACS Synthetic Biology (publishedVersion, cc-by-nc-nd)
- PubMed Central (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- Funders
- Biological and Environmental Research (DE-SC0018368, DE-SC0019404)
- 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
- Synechococcus sp. PCC 7002, DNA methylation, methyltransferase, transformation efficiency, restriction-modification systems, genome editing, metabolic engineering, halotolerant cyanobacteria
- MeSH terms
- Gene Editing, DNA, Bacterial, Escherichia coli, Methyltransferases, Transformation, Bacterial, DNA Methylation, Synechococcus, Metabolic Engineering
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