Cpf1 Is a Versatile Tool for CRISPR Genome Editing Across Diverse Species of Cyanobacteria
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directly into the desired end product using solar energy. Unfortunately, the engineering of cyanobacteria to create efficient cell factories has been impaired by the cumbersome genetic tools that are currently available for these organisms; especially when trying to accumulate multiple modifications. We sought to construct an efficient and precise tool for generating numerous markerless modifications in cyanobacteria using CRISPR technology and the alternative nuclease, Cpf1. In this study we demonstrate rapid engineering of markerless knock-ins, knock-outs and point mutations in each of three model cyanobacteria; Synechococcus, Synechocystis and Anabaena. The markerless nature of cpf1 genome editing will allow for complex genome modification that was not possible with previously existing technology while facilitating the development of cyanobacteria as highly modified biofactories.
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Ungerer and Pakrasi successfully applied a CRISPR-Cpf1 system for targeted genomic modifications in Synechococcus elongatus UTEX 2973.
“Early demonstrations of CRISPR-mediated genome editing in cyanobacteria include the work of Ungerer and Pakrasi, who successfully applied a CRISPR–Cpf1 system for targeted genomic modifications in Synechococcus elongatus UTEX 2973 [117].”
From Cyanobacteria as a Photosynthetic Chassis for Metabolic Pathway Engineering with Heterologous Gene Expression (2026) · cites this paper as [117]
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References 39 · Cited by 308
Research works printed but not linked: 2
- Fullner K. J., Gelvin S. B. & Nester E. W. In Crown Gall: Advances in Understanding Interkingdom Gene Transfer (APS Press, 1996).
- Cobb R. E., Wang Y. & Zhao H. High-efficiency multiplex genome editing of Streptomyces species using an engineered CRISPR/Cas system. ACS Synth. Biol. 4, 723– 728 (2015). 25458909 PMC4459934
- Authors
- Ungerer, J., & Pakrasi, H. B.
- Affiliations
- Washington University in St. Louis (US)
- Corresponding author
- Pakrasi, Himadri B.
- Venue
- Scientific Reports
- Published
- 2016-12-21
- Volume
- 6
- Issue
- 1
- Pages
- 39681-39681
- ISSN
- 2045-2322 (ISSN-L)
- PMC
- PMC5175191
- OpenAlex
- W2563521043
- Cited by
- 308
- References
- 39
- Citations by year
- 2026: 19 · 2025: 31 · 2024: 32 · 2023: 28 · 2022: 38 · 2021: 46 · 2020: 35 · 2019: 30 · 2018: 43 · 2017: 6
- FWCI
- 13.69
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- Scientific Reports (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Funders
- National Science Foundation (1546840)
- 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
- CRISPR/Cas12a, CRISPR gene editing, cyanobacteria, Synechococcus, Synechocystis, Anabaena, knock-in, knockout, point mutations, biofactories
- MeSH terms
- Gene Editing, Cyanobacteria, Bacterial Proteins, Carbon Dioxide, Endonucleases, Genetic Engineering, Luminescent Proteins, Microscopy, Fluorescence, Mutation, Nitrogenase, Plasmids, Anabaena, Gene Deletion, Point Mutation, Codon, Initiator, Catalytic Domain, Synechocystis, Synechococcus, CRISPR-Cas Systems
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