Evaluation of inducible promoter–riboswitch constructs for heterologous protein expression in the cyanobacterial species Anabaena sp. PCC 7120
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Abstract Cyanobacteria are promising chassis for synthetic biology applications due to the fact that they are photosynthetic organisms capable of growing in simple, inexpensive media. Given their slower growth rate than other model organisms such as Escherichia coli and Saccharomyces cerevisiae, there are fewer synthetic biology tools and promoters available for use in model cyanobacteria. Here, we compared a small library of promoter–riboswitch constructs for synthetic biology applications in Anabaena sp. PCC 7120, a model filamentous cyanobacterium. These constructs were designed from six cyanobacterial promoters of various strengths, each paired with one of two theophylline-responsive riboswitches. The promoter–riboswitch pairs were cloned upstream of a chloramphenicol acetyltransferase (cat) gene, and CAT activity was quantified using an in vitro assay. Addition of theophylline to cultures increased the CAT activity in almost all cases, allowing inducible protein production with natively constitutive promoters. We found that riboswitch F tended to have a lower induced and uninduced production compared to riboswitch E for the weak and medium promoters, although the difference was larger for the uninduced production, in accord with previous research. The strong promoters yielded a higher baseline CAT activity than medium strength and weak promoters. In addition, we observed no appreciable difference between CAT activity measured from strong promoters cultured in uninduced and induced conditions. The results of this study add to the genetic toolbox for cyanobacteria and allow future natural product and synthetic biology researchers to choose a construct that fits their needs.
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These features have contributed to growing interest in using Anabaena sp. PCC 7120 as a chassis organism for synthetic biology and metabolic pathway engineering research.
“These features have contributed to growing interest in using Anabaena sp. PCC 7120 as a chassis organism for synthetic biology and metabolic pathway engineering research [33,75].”
From Cyanobacteria as a Photosynthetic Chassis for Metabolic Pathway Engineering with Heterologous Gene Expression (2026) · cites this paper as [75]
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Cites
Thiamine derivatives bind messenger RNAs directly to regulate bacterial gene expression
Winkler, W. C., Nahvi, A., & Breaker, R. R. (2002) · Nature · doi:10.1038/nature01145 · Cited by 1,241 · see citing works
Serganov, A., & Nudler, E. (2013) · Cell · doi:10.1016/j.cell.2012.12.024 · Cited by 999 · see citing works
[57] Chloramphenicol acetyltransferase from chloramphenicol-resistant bacteria
Shaw, W. V. (1975) · Methods in enzymology on CD-ROM/Methods in enzymology · doi:10.1016/0076-6879(75)43141-x · Cited by 804 · see citing works
Genetic Control by a Metabolite Binding mRNA
Nahvi, A., Sudarsan, N., Ebert, M. S., Zou, X., Brown, K. L., & Breaker, R. R. (2002) · Chemistry & Biology · doi:10.1016/s1074-5521(02)00224-7 · Cited by 802 · see citing works
Sensing Small Molecules by Nascent RNA
Mironov, A. S., Gusarov, I., Rafikov, R., Errais, L. L., Shatalin, K. Y., Kreneva, R. A., Perumov, D. A., & Nudler, E. (2002) · Cell · doi:10.1016/s0092-8674(02)01134-0 · Cited by 682 · see citing works
Liang, P., Liu, S., Xu, F., Jiang, S., Yan, J., He, Q., Liu, W., Lin, C., Zheng, F., Wang, X., et al. (2019) · Frontiers in Microbiology · doi:10.3389/fmicb.2019.00001 · Cited by 670 · see citing works
Cyanobacteria: an emerging source for drug discovery
Singh, R. K., Tiwari, S. P., Rai, A. K., & Mohapatra, T. M. (2011) · The Journal of Antibiotics · doi:10.1038/ja.2011.21 · Cited by 370 · see citing works
Gene-Expressing Liposomes as Synthetic Cells for Molecular Communication Studies
Rampioni, G., D'Angelo, F., Leoni, L., & Stano, P. (2019) · Frontiers in Bioengineering and Biotechnology · doi:10.3389/fbioe.2019.00001 · Cited by 233 · see citing works
Tumer, N. E., Robinson, S. J., & Haselkorn, R. (1983) · Nature · doi:10.1038/306337a0 · Cited by 228 · see citing works
New applications of synthetic biology tools for cyanobacterial metabolic engineering
Santos-Merino, M., Singh, A. K., & Ducat, D. C. (2019) · Frontiers in Bioengineering and Biotechnology · doi:10.3389/fbioe.2019.00033 · Cited by 222 · see citing works
Buikema, W. J., & Haselkorn, R. (2001) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.051624898 · Cited by 221 · see citing works
Synthetic Riboswitches That Induce Gene Expression in Diverse Bacterial Species
Topp, S., Reynoso, C. M. K., Seeliger, J. C., Goldlust, I. S., Desai, S. K., Murat, D., Shen, A., Puri, A. W., Komeili, A., Bertozzi, C. R., et al. (2010) · Applied and Environmental Microbiology · doi:10.1128/aem.01537-10 · Cited by 197 · see citing works
Broad-host-range vector system for synthetic biology and biotechnology in cyanobacteria
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., et al. (2014) · Nucleic Acids Research · doi:10.1093/nar/gku673 · Cited by 186 · see citing works
Nakahira, Y., Ogawa, A., Asano, H., Oyama, T., & Tozawa, Y. (2013) · Plant and Cell Physiology · doi:10.1093/pcp/pct115 · Cited by 143 · see citing works
Ma, A. T., Schmidt, C. M., & Golden, J. W. (2014) · Applied and Environmental Microbiology · doi:10.1128/aem.01697-14 · Cited by 126 · see citing works
Toolboxes for cyanobacteria: recent advances and future direction
Sun, T., Li, S., Song, X., Diao, J., Chen, L., & Zhang, W. (2018) · Biotechnology Advances · doi:10.1016/j.biotechadv.2018.04.007 · Cited by 116 · see citing works
Frías, J. E., Flores, E., & Herrero, A. (1997) · Journal of Bacteriology · doi:10.1128/jb.179.2.477-486.1997 · Cited by 114 · see citing works
Engineering modular ‘ON’ RNA switches using biological components
Ceres, P., Trausch, J. J., & Batey, R. (2013) · Nucleic Acids Research · doi:10.1093/nar/gkt787 · Cited by 89 · see citing works
Recent advances in synthetic biology of cyanobacteria
Sengupta, A., Pakrasi, H. B., & Wangikar, P. P. (2018) · Applied Microbiology and Biotechnology · doi:10.1007/s00253-018-9046-x · Cited by 78 · see citing works
Videau, P., Wells, K. N., Singh, A. J., Gerwick, W. H., & Philmus, B. (2016) · ACS Synthetic Biology · doi:10.1021/acssynbio.6b00038 · Cited by 66 · see citing works
Self-citationNOT Gate Genetic Circuits to Control Gene Expression in Cyanobacteria
Taton, A., Ma, A. T., Ota, M., Golden, S. S., & Golden, J. W. (2017) · ACS Synthetic Biology · doi:10.1021/acssynbio.7b00203 · Cited by 51 · see citing works
A tightly inducible riboswitch system in Synechocystis sp. PCC 6803
Ohbayashi, R., Akai, H., Yoshikawa, H., Hess, W. R., & Watanabe, S. (2016) · The Journal of General and Applied Microbiology · doi:10.2323/jgam.2016.02.002 · Cited by 48 · see citing works
Heterologous Expression of Cryptomaldamide in a Cyanobacterial Host
Taton, A., Ecker, A. K., Diaz, B., Moss, N. A., Anderson, B., Reher, R., Leão, T., Simkovsky, R., Dorrestein, P. C., Gerwick, L. G., et al. (2020) · ACS Synthetic Biology · doi:10.1021/acssynbio.0c00431 · Cited by 39 · see citing works
Engineered Production of Hapalindole Alkaloids in the Cyanobacterium Synechococcus sp. UTEX 2973
Knoot, C. J., Khatri, Y., Hohlman, R. M., Sherman, D. H., & Pakrasi, H. B. (2019) · ACS Synthetic Biology · doi:10.1021/acssynbio.9b00229 · Cited by 35 · see citing works
Chi, X., Zhang, S., Sun, H., Duan, Y., Qiao, C., Luan, G., & Lü, X. (2019) · Frontiers in Microbiology · doi:10.3389/fmicb.2019.00551 · Cited by 28 · see citing works
Videau, P., Wells, K. N., Singh, A. J., Eiting, J., Proteau, P. J., & Philmus, B. (2020) · ACS Synthetic Biology · doi:10.1021/acssynbio.9b00334 · Cited by 27 · cited 1× · see citing works
“More recently, we have utilized combinatorial biosynthesis in Anabaena 7120 to produce known and unknown compounds (7), and the Gerwick and Golden laboraotries have successfully”
MethodSelf-citationCassier‐Chauvat, C., Dive, V., & Chauvat, F. (2017) · Applied Microbiology and Biotechnology · doi:10.1007/s00253-017-8105-z · Cited by 24 · see citing works
Higo, A., Isu, A., Fukaya, Y., & Hisabori, T. (2017) · Plant and Cell Physiology · doi:10.1093/pcp/pcx163 · Cited by 18 · see citing works
Higo, A., & Ehira, S. (2019) · Applied Microbiology and Biotechnology · doi:10.1007/s00253-019-09635-z · Cited by 17 · see citing works
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Harnessing the potential: Advances in cyanobacterial natural product research and biotechnology
Baunach, M., Guljamow, A., Miguel-Gordo, M., & Dittmann, E. (2024) · Natural Product Reports · doi:10.1039/d3np00045a · Cited by 66 · see citing works
Customized molecular tools to strengthen metabolic engineering of cyanobacteria
Klähn, S., Opel, F., & Hess, W. R. (2024) · Green Carbon · doi:10.1016/j.greenca.2024.05.002 · Cited by 27 · see citing works
Zhang, T., Li, S., Chen, L., Zhang, W., & Sun, T. (2024) · ACS Sustainable Chemistry & Engineering · doi:10.1021/acssuschemeng.4c04497 · Cited by 6 · see citing works
Harnessing engineered cyanobacteria for next-generation therapeutics
Bhardwaj, A., & Mishra, A. K. (2026) · Archives of Microbiology · doi:10.1007/s00203-026-04790-0 · Cited by 2 · see citing works
Zhang, T., Li, S., Chen, L., Sun, T., & Zhang, W. (2023) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/2023.08.23.554402 · Cited by 2 · see citing works
Walshe, J., & Saha, S. K. (2026) · Current Issues in Molecular Biology · doi:10.3390/cimb48060638 · Cited by 1 · see citing works
“These features have contributed to growing interest in using Anabaena sp. PCC 7120 as a chassis organism for synthetic biology and metabolic pathway engineering research [33,75].”
Yuan, X., Zheng, T., Huang, J., Li, Z., Lan, H., Ji, B., & Gao, X. (2026) · Plant Stress · doi:10.1016/j.stress.2026.101329 · Cited by 1 · see citing works
Metabolic engineering for biohydrogen production
Abubakar, A. M., Nayem, Z., Idama, E. A., Ngulde, A. B., Laghari, M., Coto, B., & Chafika, R. (2025) · Elsevier eBooks · doi:10.1016/b978-0-443-33184-8.00028-7
Development of genetic toolboxes for a bloom-forming cyanobacterium Leptolyngbya boryana FACHB-240
Liang, R., Feng, J., Chai, Y., Xia, J., Zhu, X., Ding, M., Sun, T., & Jiang, J. (2026) · Algal Research · doi:10.1016/j.algal.2026.104782
Anabaena , a multicellular model for cyanobacterial cell biology and biotechnology
Flores, E., Luque, I., & López‐Igual, R. (2026) · Journal of Bacteriology · doi:10.1128/jb.00376-26
- Authors
- Svoboda, J., Cisneros, B., & Philmus, B.
- Affiliations
- Oregon State University (US)
- Corresponding author
- Philmus, Benjamin
- Venue
- Synthetic Biology
- Publisher
- Oxford University Press (OUP)
- Published
- 2021-09-01
- Volume
- 6
- Issue
- 1
- Pages
- ysab019-ysab019
- ISSN
- 2397-7000 (ISSN-L)
- PMC
- PMC8546608
- OpenAlex
- W3197764580
- Cited by
- 11
- References
- 29
- Citations by year
- 2026: 6 · 2025: 1 · 2024: 2 · 2023: 2
- FWCI
- 0.31
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Synthetic Biology (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion, other-oa)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Funders
- Joint Genome institute (DE-AC02-05CH11231)
- National Institute of General Medical Sciences (R15GM117541)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Keywords
- Anabaena sp. PCC 7120, cyanobacteria, inducible promoters, riboswitches, heterologous expression, chloramphenicol acetyltransferase reporter, promoter strength, synthetic biology chassis
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