Abstract
Cyanobacteria are one of the most attractive hosts for biofuel production; however, genetic approaches to regulate specific chromosomal genes in cyanobacteria remain limited. With the aim of developing a novel method to regulate chromosomal gene expression in cyanobacteria, we focused on riboregulatory technology. Riboregulators are composed of two RNA fragments whose interaction leads to target gene regulation with high specificity. In this study, we inserted a riboregulator sequence upstream of the chromosomal gene encoding AbrB-like transcriptional regulator, cyAbrB2, to investigate the utility of this tool. The inserted riboregulator was able to regulate cyabrB2 gene expression, with a high ON-OFF ratio up to approximately 50-fold. The transcription levels of several genes for which cyAbrB2 acts as a transcriptional upregulator were also decreased. Further, the cyAbrB2 expression-repressed mutant showed high glycogen accumulation, equivalent to that in the cyabrB2 deletion mutant (ΔcyabrB2). Phenotypic similarities between the cyabrB2 expression-repressed mutant and the ΔcyabrB2 mutant suggest that the riboregulator can potentially be used as a new chromosomal gene regulation tool in cyanobacteria.
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This work was supported financially by the Core Research of Evolutional Science & Technology program (CREST) from the Japan Science and Technology Agency (JST).
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All experiments were performed according to applicable national and institutional guidelines for the use of microorganisms. This study does not contain any experiments involving human or animal subjects.
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Ueno, K., Sakai, Y., Shono, C. et al. Applying a riboregulator as a new chromosomal gene regulation tool for higher glycogen production in Synechocystis sp. PCC 6803. Appl Microbiol Biotechnol 101, 8465–8474 (2017). https://doi.org/10.1007/s00253-017-8570-4
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DOI: https://doi.org/10.1007/s00253-017-8570-4