Repurposing endogenous Type I-D CRISPR-Cas system for genome editing in Synechococcus sp. PCC7002
Open accessAbstract
Synechococcus sp. PCC7002 has been considered as a photosynthetic chassis for the conversion of CO 2 into biochemicals through genetic modification. However, conventional genetic manipulation techniques prove inadequate for comprehensive genetic modifications in this strain. Here, we present the development of a genome editing tool tailored for S. PCC7002, leveraging its endogenous type I-D CRISPR-Cas system. Utilizing this novel tool, we successfully deleted the glgA1 gene and iteratively edited the genome to obtain a double mutant of glgA1 and glgA2 genes. Additionally, large DNA fragments encompassing the entire type I-A (∼14 kb) or III-B CRISPR-Cas (∼21 kb) systems were completely knocked-out in S. PCC7002 using our tool. Furthermore, the endogenous pAQ5 plasmid, approximately 38 kb in length, was successfully cured from S. PCC7002. Our work demonstrates the feasibility of harnessing the endogenous CRISPR-Cas system for genome editing in S. PCC7002, thereby enriching the genetic toolkit for this species and providing a foundation for future enhancements in its biosynthetic efficiency.
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Marker-free multiplex genome editing via optimised CRISPR-Cas12a has been demonstrated in Synechocystis sp. PCC 6803 and the endogenous Type I-D CRISPR-Cas system of Synechococcus sp. PCC 7002 has been repurposed for scarless gene deletions using native cyanobacterial machinery.
“Marker-free multiplex genome editing via optimised CRISPR-Cas12a has been demonstrated in Synechocystis sp. PCC 6803 [122] and the endogenous Type I-D CRISPR-Cas system of Synechococcus sp. PCC 7002 has been repurposed for scarless gene deletions using native cyanobacterial machinery [123].”
From Cyanobacteria as a Photosynthetic Chassis for Metabolic Pathway Engineering with Heterologous Gene Expression (2026) · cites this paper as [123]
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Cyanobacterial type I CRISPR-Cas systems: distribution, mechanisms, and genome editing applications
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“Marker-free multiplex genome editing via optimised CRISPR-Cas12a has been demonstrated in Synechocystis sp. PCC 6803 [122] and the endogenous Type I-D CRISPR-Cas system of Synechococcus sp. PCC 7002 has been repurposed for scarless gene deletions using native cyanobacterial machinery [123].”
- Authors
- Yang, S., Zhang, Y., Li, C., & Tan, X.
- Affiliations
- State Key Laboratory of Biocatalysis and Enzyme Engineering (CN)
- ORCID
- Li, Chunyan
- Venue
- Microbiological Research
- Publisher
- Elsevier BV
- Published
- 2024-11
- Volume
- 288
- Pages
- 127884
- ISSN
- 0944-5013 (ISSN-L)
- OpenAlex
- W4401945004
- Cited by
- 3
- References
- 48
- Citations by year
- 2026: 2 · 2025: 1
- FWCI
- 0.41
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- www.elsevier.com/tdm/userlicense/1.0/ (tdm)
- www.elsevier.com/legal/tdmrep-license (tdm)
- www.elsevier.com/open-access/userlicense/1.0/ (vor, 365 d)
- doi.org/10.15223/policy-017 (stm-asf)
- doi.org/10.15223/policy-037 (stm-asf)
- doi.org/10.15223/policy-012 (stm-asf)
- doi.org/10.15223/policy-029 (stm-asf)
- doi.org/10.15223/policy-004 (stm-asf)
- Locations
- Microbiological Research (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Natural Science Foundation of China (31871303)
- 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 › Genetics › Bacterial Genetics and Biotechnology
- Keywords
- genome editing
- MeSH terms
- Gene Editing, Bacterial Proteins, Plasmids, Genome, Bacterial, Gene Deletion, Synechococcus, CRISPR-Cas Systems
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