Dynamic regulation of gene expression using sucrose responsive promoters and RNA interference in Saccharomyces cerevisiae

Williams, Thomas C., Espinosa, Monica I., Nielsen, Lars K. and Vickers, Claudia E. (2015) Dynamic regulation of gene expression using sucrose responsive promoters and RNA interference in Saccharomyces cerevisiae. Microbial Cell Factories, 14 43: 1-10. doi:10.1186/s12934-015-0223-7


Author Williams, Thomas C.
Espinosa, Monica I.
Nielsen, Lars K.
Vickers, Claudia E.
Title Dynamic regulation of gene expression using sucrose responsive promoters and RNA interference in Saccharomyces cerevisiae
Formatted title
Dynamic regulation of gene expression using sucrose responsive promoters and RNA interference in Saccharomyces cerevisiae
Journal name Microbial Cell Factories   Check publisher's open access policy
ISSN 1475-2859
Publication date 2015-04-01
Year available 2015
Sub-type Article (original research)
DOI 10.1186/s12934-015-0223-7
Open Access Status DOI
Volume 14
Issue 43
Start page 1
End page 10
Total pages 10
Place of publication London, United Kingdom
Publisher BioMed Central
Collection year 2016
Language eng
Formatted abstract
Background: Engineering dynamic, environmentally- and temporally-responsive control of gene expression is one of the principle objectives in the field of synthetic biology. Dynamic regulation is desirable because many engineered functions conflict with endogenous processes which have evolved to facilitate growth and survival, and minimising conflict between growth and production phases can improve product titres in microbial cell factories. There are a limited number of mechanisms that enable dynamic regulation in yeast, and fewer still that are appropriate for application in an industrial setting.

Results: To address this problem we have identified promoters that are repressed during growth on glucose, and activated during growth on sucrose. Catabolite repression and preferential glucose utilisation allows active growth on glucose before switching to production on sucrose. Using sucrose as an activator of gene expression circumvents the need for expensive inducer compounds and enables gene expression to be triggered during growth on a fermentable, high energy-yield carbon source. The ability to fine-tune the timing and population density at which gene expression is activated from the SUC2 promoter was demonstrated by varying the ratio of glucose to sucrose in the growth medium. Finally, we demonstrated that the system could also be used to repress gene expression (a process also required for many engineering projects). We used the glucose/sucrose system to control a heterologous RNA interference module and dynamically repress the expression of a constitutively regulated GFP gene.

Conclusions: The low noise levels and high dynamic range of the SUC2 promoter make it a promising option for implementing dynamic regulation in yeast. The capacity to repress gene expression using RNA interference makes the system highly versatile, with great potential for metabolic engineering applications.
Keyword Diauxic shift
GFP
Metabolic engineering
Promoter
SUC2
Sucrose
Synthetic biology
TEF1
Yeast
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2016 Collection
Australian Institute for Bioengineering and Nanotechnology Publications
 
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