Abstract
Objectives
Developing a dynamic regulation strategy is an essential step in establishing an automatic control system for manipulating metabolic fluxes and cellular behaviors. To broaden the extent of the application, a system that can generally control any gene of interest is demanded.
Results
Through characterization and optimization, the strategy repressed the immediate expression incrementally from 0 to 90% during culturing. Moreover, by changing single base pair in the lux box of the Plux promoter, the degree of repression of the target genomic gene was tuned to a difference of 70%. This strategy is expected to control metabolic flux without disrupting cell growth.
Conclusions
We engineered bacterial small RNA to develop a pathway-independent strategy that can dynamically repress the expression of any gene at the posttranscription level.
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Abbreviations
- E. coli :
-
Escherichia coli
- AHL:
-
3-Oxohexanoylhomoserine lactone
- sRNA:
-
Small RNA
- sfGFP:
-
Super-folded green fluorescent protein
- TBS:
-
Target binding sequence
- QS:
-
Quorum sensing
- OPPS:
-
Octaprenyl pyrophosphate synthase
- GOI:
-
Gene of interest
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Acknowledgements
This work was supported by the National Natural Science Foundation of China under Grant Number 81703400 (to Dr. E. Meng). The funders had no role in study design, data collection and analysis, decision to publish or preparation of the manuscript.
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EM, D-YZ and S-HB designed the appropriate technology routes. S-HB conducted main experiments including plasmid and strain construction, shake flask fermentation and western blot experiments. EM, W-YL, C-JL and D-YZ participated in the establishment of mathematic models. EM and D-YZ supervised the project and S-HB wrote the paper. All authors read and approved the final manuscript.
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Bao, SH., Li, WY., Liu, CJ. et al. Quorum-sensing based small RNA regulation for dynamic and tuneable gene expression. Biotechnol Lett 41, 1147–1154 (2019). https://doi.org/10.1007/s10529-019-02719-w
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DOI: https://doi.org/10.1007/s10529-019-02719-w