Base editing for reprogramming cyanobacterium Synechococcus elongatus
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Abstract
Cyanobacteria can directly convert carbon dioxide (CO 2) at the atmospheric level to biofuels, value-added chemicals and food products, making them ideal candidates to alleviate global climate change. Despite decades-long pioneering successes, the development of genome-editing tools, especially the CRISPR-Cas-based approaches, seems to lag behind other microbial chassis, slowing down the innovations of cyanobacteria. Here, we adapted and tailored base editing for cyanobacteria based on the CRISPR-Cas system and deamination. We achieved precise and efficient genome editing at a single-nucleotide resolution and demonstrated multiplex base editing in the model cyanobacterium Synechococcus elongatus. By using the base-editing tool, we successfully manipulated the glycogen metabolic pathway via the introduction of premature STOP codons in the relevant genes, building engineered strains with elevated potentials to produce chemicals and food from CO 2. We present here the first report of base editing in the phylum of cyanobacteria, and a paradigm for applying CRISPR-Cas systems in bacteria. We believe that our work will accelerate the metabolic engineering and synthetic biology of cyanobacteria and drive more innovations to alleviate global climate change.
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Cytosine base editing-which achieves precise C-to-T substitutions without double-strand breaks, donor DNA, or Cas nuclease toxicity-has been demonstrated in Synechococcus elongatus and subsequently extended to Synechocystis and Anabaena with multiplex capability and integrated counter-selection for plasmid curing.
“Complementing these approaches, cytosine base editing—which achieves precise C-to-T substitutions without double-strand breaks, donor DNA, or Cas nuclease toxicity—has been demonstrated in Synechococcus elongatus [124] and subsequently extended to Synechocystis and Anabaena with multiplex capability and integrated counter-selection for plasmid curing [125].”
From Cyanobacteria as a Photosynthetic Chassis for Metabolic Pathway Engineering with Heterologous Gene Expression (2026) · cites this paper as [124]
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- Authors
- Wang, S.-Y., Li, X., Wang, S.-G., & Xia, P.-F.
- Affiliations
- Shandong University (CN)
- Corresponding author
- Xia, Peng-Fei
- ORCID
- Xia, Peng-Fei
- Venue
- Metabolic Engineering
- Publisher
- Elsevier BV
- Published
- 2023-01
- Volume
- 75
- Pages
- 91-99
- ISSN
- 1096-7176 (ISSN-L)
- OpenAlex
- W4309293924
- Cited by
- 31
- References
- 58
- Citations by year
- 2026: 9 · 2025: 6 · 2024: 9 · 2023: 7
- FWCI
- 1.80
- Type
- journal article
- Language
- en
- Open access
- green
- doi.org
- Licence
- Locations
- Metabolic Engineering (publishedVersion)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- Funders
- Shandong University
- Department of Science and Technology of Shandong Province (2022HWYQ-017)
- National Natural Science Foundation of China (U20A20146)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Photosynthetic Processes and Mechanisms
- Keywords
- base editing, Anacystis nidulans, cyanobacteria, CRISPR-Cas, genome editing, glycogen metabolism, premature stop codon, CO2 conversion, synthetic biology, biofuels
- MeSH terms
- Gene Editing, Carbon Dioxide, Synechococcus, Metabolic Networks and Pathways, Metabolic Engineering, CRISPR-Cas Systems
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