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  • Title: Development of a N-Acetylneuraminic Acid-Based Sensing and Responding Switch for Orthogonal Gene Regulation in Cyanobacterial Synechococcus Strains.
    Author: Sun X, Li S, Zhang F, Sun T, Chen L, Zhang W.
    Journal: ACS Synth Biol; 2021 Aug 20; 10(8):1920-1930. PubMed ID: 34370452.
    Abstract:
    Advances in synthetic biology have allowed photosynthetic cyanobacteria as promising "green cell factories" for sustainable production of biofuels and biochemicals. However, a limited of genetic switches developed in cyanobacteria restrict the complex and orthogonal metabolic regulation. In addition, suitable and controllable switches sensing and responding to specific inducers would allow for the separation of cellular growth and expression of exogenous genes or pathways that cause metabolic burden or toxicity. Here in this study, we developed a genetic switch repressed by NanR and induced by N-acetylneuraminic acid (Neu5Ac) in a fast-growing cyanobacterium Synechococcus elongatus UTEX 2973 along with its highly homologous strain S. elongatus PCC 7942. First, nanR from Escherichia coli and a previously optimized cognate promoter PJ23119H10 were introduced into Syn2973 to control the expression of the reporter gene lacZ encoding β-galactosidase, achieving induction with negligible leakage. Second, the switch was systemically optimized to reach ∼738-fold induction by fine-tuning the expression level of NanR and introducing additional transporter of Neu5Ac. Finally, the orthogonality between the NanR/Neu5Ac switch and theophylline-responsive riboregulator was investigated, achieving a coordinated regulation or binary regulation toward the target gene. Our work here provided a new switch for transcriptional control and orthogonal regulation strategies in cyanobacteria, which would promote the metabolic regulation for the cyanobacterial chassis in the future.
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