CRISPR/Cas9 mediated targeted mutagenesis of the fast growing cyanobacterium Synechococcus elongatus UTEX 2973
Open accessAbstract
BACKGROUND: As autotrophic prokaryotes, cyanobacteria are ideal chassis organisms for sustainable production of various useful compounds. The newly characterized cyanobacterium Synechococcus elongatus UTEX 2973 is a promising candidate for serving as a microbial cell factory because of its unusually rapid growth rate. Here, we seek to develop a genetic toolkit that enables extensive genomic engineering of Synechococcus 2973 by implementing a CRISPR/Cas9 editing system. We targeted the nblA gene because of its important role in biological response to nitrogen deprivation conditions. RESULTS: First, we determined that the Streptococcus pyogenes Cas9 enzyme is toxic in cyanobacteria, and conjugational transfer of stable, replicating constructs containing the cas9 gene resulted in lethality. However, after switching to a vector that permitted transient expression of the cas9 gene, we achieved markerless editing in 100 % of cyanobacterial exconjugants after the first patch. Moreover, we could readily cure the organisms of antibiotic resistance, resulting in a markerless deletion strain. CONCLUSIONS: High expression levels of the Cas9 protein in Synechococcus 2973 appear to be toxic and result in cell death. However, introduction of a CRISPR/Cas9 genome editing system on a plasmid backbone that leads to transient cas9 expression allowed for efficient markerless genome editing in a wild type genetic background.
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
Over the past decade, significant progress has been achieved in the development of genetic tools for cyanobacterial metabolic engineering, including improved transformation methods, modular DNA assembly systems, synthetic promoters, and CRISPR-based genome editing technologies.
“Over the past decade, significant progress has been achieved in the development of genetic tools for cyanobacterial metabolic engineering, including improved transformation methods, modular DNA assembly systems, synthetic promoters, and CRISPR-based genome editing technologies [10,16,17].”
From Cyanobacteria as a Photosynthetic Chassis for Metabolic Pathway Engineering with Heterologous Gene Expression (2026) · cites this paper as [17]
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Cites
Multiplex Genome Engineering Using CRISPR/Cas Systems
Cong, L., Ran, F. A., Cox, D., Lin, S., Barretto, R., Habib, N., Hsu, P. D., Wu, X., Jiang, W., Marraffini, L. A., et al. (2013) · Science · doi:10.1126/science.1231143 · Cited by 15,339 · see citing works
Friedberg, E. C., Walker, G. C., Siede, W., Wood, R. D., Schultz, R. A., & Ellenberger, T. (2005) · doi:10.1128/9781555816704 · Cited by 5,683 · see citing works
CRISPR-Cas systems for editing, regulating and targeting genomes
Sander, J. D., & Joung, J. K. (2014) · Nature Biotechnology · doi:10.1038/nbt.2842 · Cited by 3,153 · see citing works
CRISPR RNA Maturation by Trans-Encoded Small RNA and Host Factor RNase III
Deltcheva, E., Chylinski, K., Sharma, C. M., Gonzales, K., Chao, Y., Pirzada, Z. A., Eckert, M. R., Vogel, J., & Charpentier, E. (2011) · Nature · doi:10.1038/nature09886 · Cited by 2,563 · see citing works
CRISPR/Cas, the Immune System of Bacteria and Archaea
Horvath, P., & Barrangou, R. (2010) · Science · doi:10.1126/science.1179555 · Cited by 2,471 · see citing works
RNA-guided editing of bacterial genomes using CRISPR-Cas systems
Jiang, W., Bikard, D., Cox, D., Zhang, F., & Marraffini, L. A. (2013) · Nature Biotechnology · doi:10.1038/nbt.2508 · Cited by 2,429 · see citing works
RNA-guided genetic silencing systems in bacteria and archaea
Wiedenheft, B., Sternberg, S. H., & Doudna, J. A. (2012) · Nature · doi:10.1038/nature10886 · Cited by 1,851 · see citing works
CRISPR-Cas Systems in Bacteria and Archaea: Versatile Small RNAs for Adaptive Defense and Regulation
Bhaya, D., Davison, M., & Barrangou, R. (2011) · Annual Review of Genetics · doi:10.1146/annurev-genet-110410-132430 · Cited by 914 · see citing works
Recombinational Repair of DNA Damage in Escherichia coli and Bacteriophage λ
Kuzminov, A. (1999) · Microbiology and Molecular Biology Reviews · doi:10.1128/mmbr.63.4.751-813.1999 · Cited by 897 · see citing works
Lindberg, P., Park, S., & Melis, A. (2010) · Metabolic Engineering · doi:10.1016/j.ymben.2009.10.001 · Cited by 596 · see citing works
Cobb, R. E., Wang, Y., & Zhao, H. (2014) · ACS Synthetic Biology · doi:10.1021/sb500351f · Cited by 566 · see citing works
Self-citationFatty acid production in genetically modified cyanobacteria
Liu, X., Sheng, J., & Curtiss III, R. (2011) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1103014108 · Cited by 532 · see citing works
Applications of cyanobacteria in biotechnology
Abed, R. M. M., Dobretsov, S., & Sudesh, K. (2009) · Journal of Applied Microbiology · doi:10.1111/j.1365-2672.2008.03918.x · Cited by 528 · see citing works
Ethanol Synthesis by Genetic Engineering in Cyanobacteria
Deng, M.-D., & Coleman, J. R. (1999) · Applied and Environmental Microbiology · doi:10.1128/aem.65.2.523-528.1999 · Cited by 457 · see citing works
He, Y., Zhang, L., Zhang, J., & Bao, J. (2014) · Biotechnology for Biofuels · doi:10.1186/1754-6834-7-1 · Cited by 404 · see citing works
[12] Genetic engineering of the cyanobacterial chromosome
Golden, S. S., Brusslan, J. A., & Haselkorn, R. (1987) · Methods in enzymology on CD-ROM/Methods in enzymology · doi:10.1016/0076-6879(87)53055-5 · Cited by 395 · see citing works
Yu, J., Liberton, M., Cliften, P. F., Head, R. D., Jacobs, J., Smith, R. D., Koppenaal, D., Brand, J. J., & Pakrasi, H. B. (2015) · Scientific Reports · doi:10.1038/srep08132 · Cited by 372 · see citing works
Self-citationElucidation of the molecular structures of components of the phycobilisome: reconstructing a giant
Adir, N. (2005) · Photosynthesis Research · doi:10.1007/s11120-004-2143-y · Cited by 340 · see citing works
Wolk, C. P., Vonshak, A., Kehoe, P., & Elhai, J. (1984) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.81.5.1561 · Cited by 328 · see citing works
The cyanobacterial genome core and the origin of photosynthesis
Mulkidjanian, A. Y., Koonin, E. V., Makarova, K. S., Mekhedov, S. L., Sorokin, A. V., Wolf, Y. I., Dufresne, A., Partensky, F., Burd, H., Kaznadzey, D., et al. (2006) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0605709103 · Cited by 325 · see citing works
Chen, X., Shekiro, J., Pschorn, T., Sabourin, M., Tao, L., Elander, R., Park, S., Jennings, E., Nelson, R., Trass, O., et al. (2014) · Biotechnology for Biofuels · doi:10.1186/1754-6834-7-98 · Cited by 300 · see citing works
Collier, J. L., & Grossman, A. R. (1994) · The EMBO Journal · doi:10.1002/j.1460-2075.1994.tb06352.x · Cited by 271 · see citing works
Griese, M., Lange, C., & Soppa, J. (2011) · FEMS Microbiology Letters · doi:10.1111/j.1574-6968.2011.02368.x · Cited by 264 · see citing works
Multiple Gene Repression in Cyanobacteria Using CRISPRi
Yao, L., Cengic, I., Anfelt, J., & Hudson, E. P. (2016) · ACS Synthetic Biology · doi:10.1021/acssynbio.5b00264 · Cited by 246 · see citing works
Efficient Genome Editing in Clostridium cellulolyticum via CRISPR-Cas9 Nickase
Xu, T., Li, Y., Shi, Z. J., Hemme, C. L., Li, Y., Zhu, Y., Nostrand, J. D. V., He, Z., & Zhou, J. Z. (2015) · Applied and Environmental Microbiology · doi:10.1128/aem.00873-15 · Cited by 223 · see citing works
Muth, G., Nußbaumer, B., Wohlleben, W., & Pühler, A. (1989) · Molecular and General Genetics MGG · doi:10.1007/bf00259605 · Cited by 222 · see citing works
McNeely, K., Xu, Y., Bennette, N., Bryant, D. A., & Dismukes, G. C. (2010) · Applied and Environmental Microbiology · doi:10.1128/aem.00862-10 · Cited by 168 · see citing works
Heidelberg, J. F., Nelson, W. C., Schoenfeld, T. W., & Bhaya, D. (2009) · PLoS ONE · doi:10.1371/journal.pone.0004169 · Cited by 167 · see citing works
The Responses of Cyanobacteria to Environmental Conditions: Light and Nutrients
Grossman, A. R., Schaefer, M. R., Chiang, G. G., & Collier, J. L. (1994) · Springer eBooks · doi:10.1007/978-94-011-0227-8_21 · Cited by 137 · see citing works
Exploring the photosynthetic production capacity of sucrose by cyanobacteria
Du, W., Liang, F., Duan, Y., Tan, X., & Lü, X. (2013) · Metabolic Engineering · doi:10.1016/j.ymben.2013.05.001 · Cited by 133 · see citing works
Scholz, I., Lange, S. J., Hein, S., Hess, W. R., & Backofen, R. (2013) · PLoS ONE · doi:10.1371/journal.pone.0056470 · Cited by 128 · see citing works
Evidence for the widespread distribution of CRISPR-Cas system in the PhylumCyanobacteria
Cai, F., Axen, S. D., & Kerfeld, C. A. (2013) · RNA Biology · doi:10.4161/rna.24571 · Cited by 98 · see citing works
Sakai, M., Ogawa, T., Matsuoka, M., & Fukuda, H. (1997) · Journal of Fermentation and Bioengineering · doi:10.1016/s0922-338x(97)82004-1 · Cited by 89 · see citing works
Marraccini, P., Bulteau, S., Cassier‐Chauvat, C., Mermet-Bouvier, P., & Chauvat, F. (1993) · Plant Molecular Biology · doi:10.1007/bf00021546 · Cited by 87 · see citing works
Improved production of fatty alcohols in cyanobacteria by metabolic engineering
Yao, L., Qi, F., Tan, X., & Lü, X. (2014) · Biotechnology for Biofuels · doi:10.1186/1754-6834-7-94 · Cited by 87 · see citing works
Vectors for the complementation analysis of cyanobacterial mutants
Зинченко, В. В., Piven, I., Мельник, В. А., & Shestakov, S. V. (1999) · Russian Journal of Genetics · Cited by 86 · see citing works
Biotechnological uses of cyanobacteria
Lem, N. W., & Glick, B. R. (1985) · Biotechnology Advances · doi:10.1016/0734-9750(85)90291-5 · Cited by 48 · see citing works
Matsuoka, M., Takahama, K., & Ogawa, T. (2001) · Microbiology · doi:10.1099/00221287-147-8-2077 · Cited by 37 · see citing works
Flagged first, then by citations
Cited by
Sorted by citations
Calero, P., & Nikel, P. I. (2018) · Microbial Biotechnology · doi:10.1111/1751-7915.13292 · Cited by 324 · see citing works
Cpf1 Is a Versatile Tool for CRISPR Genome Editing Across Diverse Species of Cyanobacteria
Ungerer, J., & Pakrasi, H. B. (2016) · Scientific Reports · doi:10.1038/srep39681 · Cited by 308 · see citing works
Self-citationRecent Developments on Genetic Engineering of Microalgae for Biofuels and Bio‐Based Chemicals
Ng, I., Tan, S., Kao, P., Chang, Y.-K., & Chang, J. (2017) · Biotechnology Journal · doi:10.1002/biot.201600644 · Cited by 253 · see citing works
CRISPR-Cas9 for the genome engineering of cyanobacteria and succinate production
Li, H., Shen, C. R., Huang, C.-H., Sung, L., Wu, M., & Hu, Y. (2016) · Metabolic Engineering · doi:10.1016/j.ymben.2016.09.006 · Cited by 232 · 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
Development of a confinable gene drive system in the human disease vector Aedes aegypti
Li, M., Yang, T., Kandul, N. P., Bui, M., Gamez, S., Raban, R. R., Bennett, J. B., C., H. M. S., Lanzaro, G. C., Schmidt, H., et al. (2020) · eLife · doi:10.7554/elife.51701 · Cited by 217 · see citing works
Genetic engineering of microalgae for enhanced biorefinery capabilities
Fayyaz, M., Chew, K. W., Show, P. L., Ling, T. C., Ng, I.-S., & Chang, J.-S. (2020) · Biotechnology Advances · doi:10.1016/j.biotechadv.2020.107554 · Cited by 225 · see citing works
Application of different types of CRISPR/Cas-based systems in bacteria
Liu, Z., Dong, H., Cui, Y., Cong, L., & Zhang, D. (2020) · Microbial Cell Factories · doi:10.1186/s12934-020-01431-z · Cited by 204 · see citing works
Cyanogate: a modular cloning suite for engineering cyanobacteria based on the plant moclo syntax
Vasudevan, R., Gale, G. A. R., Schiavon, A. A., Puzorjov, A., Malin, J., Gillespie, M. D., Vavitsas, K., Zulkower, V., Wang, B., Howe, C. J., et al. (2019) · Plant Physiology · doi:10.1104/pp.18.01401 · Cited by 189 · see citing works
Current status and perspectives of genome editing technology for microalgae
Jeon, S., Lim, J.-M., Lee, H.-G., Shin, S.-E., Kang, N. K., Park, Y.-I., Oh, H.-M., Jeong, W.-J., Jeong, B.-R., & Chang, Y. K. (2017) · Biotechnology for Biofuels · doi:10.1186/s13068-017-0957-z · Cited by 154 · see citing works
CRISPR-Cas9/Cas12a biotechnology and application in bacteria
Yao, R., Liu, D., Jia, X., Zheng, Y., Liu, W. Y., & Xiao, Y. (2018) · Synthetic and Systems Biotechnology · doi:10.1016/j.synbio.2018.09.004 · Cited by 159 · see citing works
CRISPR technologies for bacterial systems: Current achievements and future directions
Choi, K. R., & Lee, S. Y. (2016) · Biotechnology Advances · doi:10.1016/j.biotechadv.2016.08.002 · Cited by 147 · see citing works
Development of a CRISPR/Cas9 genome editing toolbox for Corynebacterium glutamicum
Liu, J., Wang, Y., Lu, Y., Zheng, P., Sun, J., & Ma, Y. (2017) · Microbial Cell Factories · doi:10.1186/s12934-017-0815-5 · Cited by 143 · see citing works
Engineering Halomonas species TD01 for enhanced polyhydroxyalkanoates synthesis via CRISPRi
Wei, T., Lv, L., & Chen, G. (2017) · Microbial Cell Factories · doi:10.1186/s12934-017-0655-3 · Cited by 141 · see citing works
CRISPR-Based Technologies for Metabolic Engineering in Cyanobacteria
Behler, J., Vijay, D., Hess, W. R., & Akhtar, M. K. (2018) · Trends in biotechnology · doi:10.1016/j.tibtech.2018.05.011 · Cited by 138 · see citing works
Microalgae-based biotechnological sequestration of carbon dioxide for net zero emissions
Ma, Z., Cheah, W. Y., Ng, I., Chang, J., Zhao, M., & Show, P. L. (2022) · Trends in biotechnology · doi:10.1016/j.tibtech.2022.09.002 · Cited by 138 · see citing works
Ng, I., Keskin, B. B., & Tan, S. (2020) · Biotechnology Journal · doi:10.1002/biot.201900228 · Cited by 135 · see citing works
CRISPR/Cas-based genome engineering in natural product discovery
Tong, Y., Weber, T., & Lee, S. Y. (2018) · Natural Product Reports · doi:10.1039/c8np00089a · Cited by 130 · see citing works
Development of CRISPR/Cas9 system in Chlorella vulgaris FSP-E to enhance lipid accumulation
Lin, W., & Ng, I. (2019) · Enzyme and Microbial Technology · doi:10.1016/j.enzmictec.2019.109458 · Cited by 121 · see citing works
Niu, T., Lin, G., Xie, L.-R., Wang, Z.-Q., Xing, W., Zhang, J., & Zhang, C. (2018) · ACS Synthetic Biology · doi:10.1021/acssynbio.8b00437 · Cited by 119 · 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
Joyia, M. A. K., Ahmad, M., Chen, Y.-F., Mustaqeem, M., Ali, A., Abbas, A., & Gondal, M. A. (2024) · Energy Conversion and Management · doi:10.1016/j.enconman.2024.119037 · Cited by 117 · see citing works
Li, S., Sun, T., Xu, C., Chen, L., & Zhang, W. (2018) · Metabolic Engineering · doi:10.1016/j.ymben.2018.06.002 · Cited by 112 · see citing works
Engineering cyanobacteria for production of terpenoids
Lin, P.-C., & Pakrasi, H. B. (2018) · Planta · doi:10.1007/s00425-018-3047-y · Cited by 109 · see citing works
Self-citationCyanobacteria as photoautotrophic biofactories of high-value chemicals
Noreña-Caro, D., & Benton, M. G. (2018) · Journal of CO2 Utilization · doi:10.1016/j.jcou.2018.10.008 · Cited by 109 · see citing works
Lin, W., Tan, S., Hsiang, C., Sung, P.-K., & Ng, I. (2019) · Bioresource Technology · doi:10.1016/j.biortech.2019.121932 · Cited by 108 · see citing works
Metabolic Engineering of Oleaginous Yeasts for Production of Fuels and Chemicals
Shi, S., & Zhao, H. (2017) · Frontiers in Microbiology · doi:10.3389/fmicb.2017.02185 · Cited by 107 · see citing works
Self-citationAdvances in metabolic engineering in the microbial production of fuels and chemicals from C1 gas
Humphreys, C. M., & Minton, N. P. (2018) · Current Opinion in Biotechnology · doi:10.1016/j.copbio.2017.12.023 · Cited by 104 · see citing works
Advances and challenges in genetic engineering of microalgae
Fajardo, C., Donato, M. D., Carrasco-Reinado, R., Martı́nez-Rodrı́guez, G., Mancera, J. M., & Fernández‐Acero, F. J. (2019) · Reviews in Aquaculture · doi:10.1111/raq.12322 · Cited by 103 · see citing works
Systems and synthetic biology for the biotechnological application of cyanobacteria
Hagemann, M., & Hess, W. R. (2017) · Current Opinion in Biotechnology · doi:10.1016/j.copbio.2017.07.008 · Cited by 101 · see citing works
Wang, Q., & Coleman, J. J. (2019) · Computational and Structural Biotechnology Journal · doi:10.1016/j.csbj.2019.06.007 · Cited by 97 · see citing works
Zhu, H., & Liang, C. (2019) · Bioinformatics · doi:10.1093/bioinformatics/bty1061 · Cited by 86 · see citing works
Shabestary, K., & Hudson, E. P. (2016) · Metabolic Engineering Communications · doi:10.1016/j.meteno.2016.07.003 · Cited by 90 · see citing works
Editing streptomycete genomes in the CRISPR/Cas9 age
Alberti, F., & Corre, C. (2019) · Natural Product Reports · doi:10.1039/c8np00081f · Cited by 90 · see citing works
Cyanobacteria as Chassis for Industrial Biotechnology: Progress and Prospects
Al‐Haj, L., Lui, Y. T., Abed, R. M. M., Gomaa, M., & Purton, S. (2016) · Life · doi:10.3390/life6040042 · Cited by 90 · see citing works
Comparative Genomics of DNA Recombination and Repair in Cyanobacteria: Biotechnological Implications
Cassier‐Chauvat, C., Veaudor, T., & Chauvat, F. (2016) · Frontiers in Microbiology · doi:10.3389/fmicb.2016.01809 · Cited by 86 · see citing works
Cas9 off-target binding to the promoter of bacterial genes leads to silencing and toxicity
Rostain, W., Grebert, T., Vyhovskyi, D., Pizarro, P. T., Tshinsele-Van Bellingen, G., Cui, L., & Bikard, D. (2023) · Nucleic Acids Research · doi:10.1093/nar/gkad170 · Cited by 85 · see citing works
CRISPR Tools To Control Gene Expression in Bacteria
Vigouroux, A., & Bikard, D. (2020) · Microbiology and Molecular Biology Reviews · doi:10.1128/mmbr.00077-19 · Cited by 82 · 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
Self-citationApplication of the CRISPR/Cas system for genome editing in microalgae
Zhang, Y., Jiang, J., Shi, T., Sun, X., Zhao, Q., Huang, H., & Ren, L. (2019) · Applied Microbiology and Biotechnology · doi:10.1007/s00253-019-09726-x · Cited by 80 · see citing works
Development and Application of CRISPR/Cas in Microbial Biotechnology
Ding, W., Zhang, Y., & Shi, S. (2020) · Frontiers in Bioengineering and Biotechnology · doi:10.3389/fbioe.2020.00711 · Cited by 78 · see citing works
Solar-to-chemical and solar-to-fuel production from CO2 by metabolically engineered microorganisms
Woo, H. M. (2017) · Current Opinion in Biotechnology · doi:10.1016/j.copbio.2016.11.017 · Cited by 76 · see citing works
Choi, S. Y., & Woo, H. M. (2020) · ACS Synthetic Biology · doi:10.1021/acssynbio.0c00091 · Cited by 76 · see citing works
Metabolic engineering tools in model cyanobacteria
Carroll, A. L., Case, A. E., Zhang, A., & Atsumi, S. (2018) · Metabolic Engineering · doi:10.1016/j.ymben.2018.03.014 · Cited by 75 · see citing works
A CRISPR‐Cpf1‐Assisted Non‐Homologous End Joining Genome Editing System of Mycobacterium smegmatis
Sun, B., Yang, J., Yang, S., Ye, R. D., Chen, D., & Jiang, Y. (2018) · Biotechnology Journal · doi:10.1002/biot.201700588 · Cited by 75 · see citing works
Godbole, V., Pal, M. K., & Gautam, P. (2021) · Algal Research · doi:10.1016/j.algal.2021.102436 · Cited by 74 · see citing works
CRISPR-based metabolic pathway engineering
Zhao, D., Zhu, X., Zhou, H., Sun, N., Wang, T., Bi, C., & Zhang, X. (2020) · Metabolic Engineering · doi:10.1016/j.ymben.2020.10.004 · Cited by 70 · see citing works
Lee, H. J., Lee, J., Lee, S., Um, Y., Kim, Y., Sim, S. J., Choi, J., & Woo, H. M. (2017) · Journal of Agricultural and Food Chemistry · doi:10.1021/acs.jafc.7b03625 · Cited by 69 · see citing works
Progress and challenges in engineering cyanobacteria as chassis for light‐driven biotechnology
Hitchcock, A., Hunter, C. N., & Canniffe, D. P. (2019) · Microbial Biotechnology · doi:10.1111/1751-7915.13526 · Cited by 66 · see citing works
Progress of CRISPR‐Cas Based Genome Editing in Photosynthetic Microbes
Naduthodi, M. I. S., Barbosa, M. J., & John van der, O. O. S. T. (2018) · Biotechnology Journal · doi:10.1002/biot.201700591 · Cited by 63 · see citing works
Biocatalysis in Green and Blue: Cyanobacteria
Jodlbauer, J., Rohr, T., Oliver Spadiut, A., Mihovilovič, M. D., & Rudroff, F. (2021) · Trends in biotechnology · doi:10.1016/j.tibtech.2020.12.009 · Cited by 63 · see citing works
Emerging Species and Genome Editing Tools: Future Prospects in Cyanobacterial Synthetic Biology
Gale, G. A. R., Schiavon Osorio, A. A., Mills, L. A., Wang, B., Lea-Smith, D. J., & McCormick, A. J. (2019) · Microorganisms · doi:10.3390/microorganisms7100409 · Cited by 61 · see citing works
“Synechococcus elongatus UTEX 2973 SpCas9 episomal [58]”
BackgroundInfluentialGenome-Scale Fluxome of Synechococcus elongatus UTEX 2973 Using Transient 13 C-Labeling Data
Hendry, J. I., Gopalakrishnan, S., Ungerer, J. L., Pakrasi, H. B., Tang, Y., & Maranas, C. D. (2018) · PLANT PHYSIOLOGY · doi:10.1104/pp.18.01357 · Cited by 61 · see citing works
Self-citationOptimizing Systems for Cas9 Expression in Toxoplasma gondii
Markus, B. M., Bell, G. W., Lorenzi, H., & Lourido, S. (2019) · mSphere · doi:10.1128/msphere.00386-19 · Cited by 58 · see citing works
A Novel Biocontainment Strategy Makes Bacterial Growth and Survival Dependent on Phosphite
Hirota, R., Abe, K., Katsuura, Z.-I., Noguchi, R., Moribe, S., Motomura, K., Ishida, T., Alexandrov, M., Funabashi, H., Ikeda, T., et al. (2017) · Scientific Reports · doi:10.1038/srep44748 · Cited by 59 · see citing works
Knoot, C. J., Biswas, S., & Pakrasi, H. B. (2019) · ACS Synthetic Biology · doi:10.1021/acssynbio.9b00417 · Cited by 59 · see citing works
Self-citationIdentifying the Metabolic Differences of a Fast-Growth Phenotype in Synechococcus UTEX 2973
Mueller, T. J., Ungerer, J. L., Pakrasi, H. B., & Maranas, C. D. (2017) · Scientific Reports · doi:10.1038/srep41569 · Cited by 58 · see citing works
Self-citationSynthetic Biology Toolkits for Metabolic Engineering of Cyanobacteria
Xia, P., Ling, H., Foo, J. L., & Chang, M. W. (2019) · Biotechnology Journal · doi:10.1002/biot.201800496 · Cited by 58 · see citing works
Carbon-negative synthetic biology: challenges and emerging trends of cyanobacterial technology
Tan, C., Xu, P., & Tao, F. (2022) · Trends in biotechnology · doi:10.1016/j.tibtech.2022.09.012 · Cited by 55 · see citing works
Biology and applications of CRISPR–Cas12 and transposon-associated homologs
Wu, W. Y., Adiego-Pérez, B., & John van der, O. O. S. T. (2024) · Nature Biotechnology · doi:10.1038/s41587-024-02485-9 · Cited by 54 · see citing works
Emerging molecular biology tools and strategies for engineering natural product biosynthesis
Xu, W., Klumbys, E., Ang, E. L., & Zhao, H. (2019) · Metabolic Engineering Communications · doi:10.1016/j.mec.2019.e00108 · Cited by 55 · see citing works
Self-citationSengupta, A., Pritam, P., Jaiswal, D., Bandyopadhyay, A., Pakrasi, H. B., & Wangikar, P. P. (2020) · Metabolites · doi:10.3390/metabo10060250 · Cited by 52 · see citing works
Self-citationRegulatory systems for gene expression control in cyanobacteria
Till, P., Toepel, J., Bühler, B., Mach, R. L., & Mach-Aigner, A. R. (2020) · Applied Microbiology and Biotechnology · doi:10.1007/s00253-019-10344-w · Cited by 54 · see citing works
Feng, S., Xie, X., Liu, J., Li, A., Wang, Q., Guo, D., Li, S., Li, Y., Wang, Z., Guo, T., et al. (2023) · Journal of Nanobiotechnology · doi:10.1186/s12951-023-02139-z · Cited by 58 · see citing works
The primary transcriptome of the fast-growing cyanobacterium Synechococcus elongatus UTEX 2973
Tan, X., Hou, S., Song, K., Georg, J., Klähn, S., Lü, X., & Hess, W. R. (2018) · Biotechnology for Biofuels · doi:10.1186/s13068-018-1215-8 · Cited by 54 · see citing works
Targeted Nucleotide Editing Technologies for Microbial Metabolic Engineering
Arazoe, T., Kondo, A., & Nishida, K. (2018) · Biotechnology Journal · doi:10.1002/biot.201700596 · Cited by 54 · see citing works
Recent Progress and Challenges in CRISPR-Cas9 Engineered Algae and Cyanobacteria
Patel, V. K., Das, A., Kumari, R., & Kajla, S. (2023) · Algal Research · doi:10.1016/j.algal.2023.103068 · Cited by 50 · see citing works
Current Metabolic Engineering Strategies for Photosynthetic Bioproduction in Cyanobacteria
Satta, A., Esquirol, L., & Ebert, B. E. (2023) · Microorganisms · doi:10.3390/microorganisms11020455 · Cited by 51 · see citing works
The Synthetic Biology Toolkit for Photosynthetic Microorganisms
Vavitsas, K., Crozet, P., Vinde, M. H., Davies, F. K., Lemaire, S. D., & Vickers, C. E. (2019) · PLANT PHYSIOLOGY · doi:10.1104/pp.19.00345 · Cited by 50 · see citing works
Recent advances in synthetic biology of cyanobacteria for improved chemicals production
Wang, F., Gao, Y., & Yang, G. (2020) · Bioengineered · doi:10.1080/21655979.2020.1837458 · Cited by 49 · see citing works
Recent advances of Cas12a applications in bacteria
Meliawati, M., Schilling, C., & Schmid, J. (2021) · Applied Microbiology and Biotechnology · doi:10.1007/s00253-021-11243-9 · Cited by 49 · see citing works
Lee, T., Lin, J., Tsai, T.-H., Yang, R., & Ng, I. (2022) · Bioresource Technology · doi:10.1016/j.biortech.2022.128350 · Cited by 49 · see citing works
Efficient Cas9-based genome editing of Rhodobacter sphaeroides for metabolic engineering
Mougiakos, I., Orsi, E., Ghiffary, M. R., Post, W. B., Maria, A. D., Adiego-Pérez, B., Kengen, S. W. M., Weusthuis, R. A., & John van der, O. O. S. T. (2019) · Microbial Cell Factories · doi:10.1186/s12934-019-1255-1 · Cited by 44 · see citing works
Cui, J., Sun, H., Chen, R., Sun, J., Mo, G., Luan, G., & Lü, X. (2023) · Green Carbon · doi:10.1016/j.greenca.2023.11.004 · Cited by 48 · see citing works
Cui, J., Sun, T., Li, S., Xie, Y., Song, X., Wang, F., Chen, L., & Zhang, W. (2020) · Frontiers in Bioengineering and Biotechnology · doi:10.3389/fbioe.2020.00500 · Cited by 46 · see citing works
Engineering salt tolerance of photosynthetic cyanobacteria for seawater utilization
Cui, J., Sun, T., Chen, L., & Zhang, W. (2020) · Biotechnology Advances · doi:10.1016/j.biotechadv.2020.107578 · Cited by 44 · see citing works
Photosynthetic CO2 Conversion to Fatty Acid Ethyl Esters (FAEEs) Using Engineered Cyanobacteria
Lee, H. J., Choi, J., Lee, S., Um, Y., Sim, S. J., Kim, Y., & Woo, H. M. (2017) · Journal of Agricultural and Food Chemistry · doi:10.1021/acs.jafc.7b00002 · Cited by 46 · see citing works
Pattharaprachayakul, N., Lee, M., Incharoensakdi, A., & Woo, H. M. (2020) · Enzyme and Microbial Technology · doi:10.1016/j.enzmictec.2020.109619 · Cited by 43 · see citing works
A Cloning-Free Method for CRISPR/Cas9-Mediated Genome Editing in Fission Yeast
Zhang, X., He, J.-B., Wang, Y.-Z., & Du, L. (2018) · G3 Genes Genomes Genetics · doi:10.1534/g3.118.200164 · Cited by 43 · see citing works
SEVA-Cpf1, a CRISPR-Cas12a vector for genome editing in cyanobacteria
Baldanta, S., Guevara, G., & Navarro-Llorens, J. M. (2022) · Microbial Cell Factories · doi:10.1186/s12934-022-01830-4 · Cited by 42 · see citing works
Ajeng, A. A., Rosli, N. S. M., Abdullah, R., Yaacob, J. S., Qi, N. C., & Loke, S. P. (2022) · Journal of Biotechnology · doi:10.1016/j.jbiotec.2022.10.011 · Cited by 39 · see citing works
Regulatory Tools for Controlling Gene Expression in Cyanobacteria
Gordon, G. C., & Pfleger, B. F. (2018) · Advances in experimental medicine and biology · doi:10.1007/978-981-13-0854-3_12 · Cited by 40 · see citing works
Muthukrishnan, L. (2022) · Journal of Basic Microbiology · doi:10.1002/jobm.202100417 · Cited by 40 · see citing works
Liu, D., Johnson, V. M., & Pakrasi, H. B. (2020) · ACS Synthetic Biology · doi:10.1021/acssynbio.0c00106 · Cited by 37 · see citing works
Self-citationDoing synthetic biology with photosynthetic microorganisms
Vavitsas, K., Kugler, A., Satta, A., Hatzinikolaou, D. G., Lindblad, P. C., Fewer, D. P., Lindberg, P., Toivari, M. H., & Stensjö, K. (2021) · Physiologia Plantarum · doi:10.1111/ppl.13455 · Cited by 39 · see citing works
Natural transformation of the filamentous cyanobacterium Phormidium lacuna
Nies, F., Mielke, M., Pochert, J., & Lamparter, T. (2020) · PLoS ONE · doi:10.1371/journal.pone.0234440 · Cited by 37 · see citing works
CRISPR‐Enabled Tools for Engineering Microbial Genomes and Phenotypes
Tarasava, K., Oh, E. J., Eckert, C. A., & Gill, R. T. (2018) · Biotechnology Journal · doi:10.1002/biot.201700586 · Cited by 37 · see citing works
Characterization of Cas proteins for CRISPR‐Cas editing in streptomycetes
Yeo, W. L., Heng, E., Tan, L. L., Lim, Y. W., Lim, Y. H., Hoon, S. S., Zhao, H., Zhang, M. M., & Wong, F. T. (2019) · Biotechnology and Bioengineering · doi:10.1002/bit.27021 · Cited by 37 · see citing works
Self-citationDeveloping a Cas9‐based tool to engineer native plasmids in Synechocystis sp. PCC 6803
Xiao, Y., Wang, S., Rommelfanger, S. R., Balassy, A., Barba‐Ostria, C., Gu, P., Galazka, J. M., & Zhang, F. (2018) · Biotechnology and Bioengineering · doi:10.1002/bit.26747 · Cited by 36 · see citing works
Khan, A. Z., Bilal, M., Mehmood, S., Sharma, A. K., & Iqbal, H. M. N. (2019) · Life · doi:10.3390/life9030054 · Cited by 36 · see citing works
Verma, S., Thapa, S., Siddiqui, N. M., & Chakdar, H. (2022) · World Journal of Microbiology and Biotechnology · doi:10.1007/s11274-022-03285-6 · Cited by 36 · see citing works
Cengic, I., Cañadas, I. C., Minton, N. P., & Hudson, E. P. (2022) · ACS Synthetic Biology · doi:10.1021/acssynbio.2c00375 · Cited by 36 · see citing works
Czajka, J. J., Wang, Q., Wang, Y., & Tang, Y. (2017) · Applied Microbiology and Biotechnology · doi:10.1007/s00253-017-8489-9 · Cited by 35 · 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
Self-citationHeterologous production of cyanobacterial compounds
Dhakal, D., Chen, M., Luesch, H., & Ding, Y. (2021) · Journal of Industrial Microbiology and Biotechnology · doi:10.1093/jimb/kuab003 · Cited by 33 · see citing works
Krumbach, K., Sonntag, C. K., Eggeling, L., & Marienhagen, J. (2019) · ACS Synthetic Biology · doi:10.1021/acssynbio.9b00361 · Cited by 33 · see citing works
Synthetic Biology Toolbox for Nitrogen-Fixing Soil Microbes
Venkataraman, M., Yñigez‐Gutierrez, A. E., Infante, V., MacIntyre, A. M., Fernandes‐Júnior, P. I., Ané, J., & Pfleger, B. F. (2023) · ACS Synthetic Biology · doi:10.1021/acssynbio.3c00414 · Cited by 33 · see citing works
Knoot, C. J., & Pakrasi, H. B. (2019) · Scientific Reports · doi:10.1038/s41598-018-38124-y · Cited by 32 · see citing works
Self-citationAdvancing the design and delivery of CRISPR antimicrobials
Fagen, J. R., Collias, D., Singh, A. K., & Beisel, C. L. (2017) · Current Opinion in Biomedical Engineering · doi:10.1016/j.cobme.2017.10.001 · Cited by 32 · see citing works
Base editing for reprogramming cyanobacterium Synechococcus elongatus
Wang, S.-Y., Li, X., Wang, S.-G., & Xia, P.-F. (2023) · Metabolic Engineering · doi:10.1016/j.ymben.2022.11.005 · Cited by 31 · see citing works
- Authors
- Wendt, K. E., Ungerer, J., Cobb, R. E., Zhao, H., & Pakrasi, H. B.
- Affiliations
- Washington University in St. Louis (US)
- University of Illinois Urbana-Champaign (US)
- Corresponding author
- Pakrasi, Himadri B.
- Venue
- Microbial Cell Factories
- Published
- 2016-06-23
- Volume
- 15
- Issue
- 1
- Pages
- 115-115
- ISSN
- 1475-2859 (ISSN-L)
- PMC
- PMC4917971
- OpenAlex
- W2470889463
- Cited by
- 232
- References
- 38
- Citations by year
- 2026: 15 · 2025: 15 · 2024: 24 · 2023: 17 · 2022: 29 · 2021: 23 · 2020: 22 · 2019: 34 · 2018: 31 · 2017: 16 · 2016: 6
- FWCI
- 15.07
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- Microbial Cell Factories (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- OSTI OAI (U.S. Department of Energy Office of Scientific and Technical Information) (submittedVersion)
- Funders
- Office of Science
- National Institute of General Medical Sciences (GM077596)
- 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/Cas9, cyanobacteria, genome editing, conjugative transfer
- MeSH terms
- Endonucleases, Genetic Techniques, Mutagenesis, Synechococcus, Clustered Regularly Interspaced Short Palindromic Repeats
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
