Synthetic gene circuit-mediated monitoring of endogenous metabolites

Identification of GAL11 as a novel multicopy enhancer of S-adenosylmethionine level in yeast

Taichi Umeyama, Satoshi Okada, Takashi Ito

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Monitoring levels of key metabolites in living cells comprises a critical step in various investigations. The simplest approach to this goal is a fluorescent reporter gene using an endogenous promoter responsive to the metabolite. However, such a promoter is often not identified or even present in the species of interest. An alternative can be a synthetic gene circuit based on a heterologous pair consisting of a promoter and a transcription factor known to respond to the metabolite. We exploited the met operator and MetJ repressor of Escherichia coli, the interaction between which depends on S-adenosylmethionine (SAM), to construct synthetic gene circuits that report SAM levels in Saccharomyces cerevisiae. Using a dual-input circuit that outputs selection marker genes in a doxycycline-tunable manner, we screened a genomic library to identify GAL11 as a novel multicopy enhancer of SAM levels. These results demonstrate the potential and utility of synthetic gene circuit-mediated metabolite monitoring.

Original languageEnglish
Pages (from-to)425-430
Number of pages6
JournalACS Synthetic Biology
Volume2
Issue number8
DOIs
Publication statusPublished - Aug 16 2013
Externally publishedYes

Fingerprint

Synthetic Genes
S-Adenosylmethionine
Gene Regulatory Networks
Metabolites
Yeast
Genes
Yeasts
Networks (circuits)
Monitoring
Genomic Library
Doxycycline
Reporter Genes
Saccharomyces cerevisiae
Transcription Factors
Transcription factors
Escherichia coli
Cells

All Science Journal Classification (ASJC) codes

  • Biomedical Engineering
  • Biochemistry, Genetics and Molecular Biology (miscellaneous)

Cite this

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abstract = "Monitoring levels of key metabolites in living cells comprises a critical step in various investigations. The simplest approach to this goal is a fluorescent reporter gene using an endogenous promoter responsive to the metabolite. However, such a promoter is often not identified or even present in the species of interest. An alternative can be a synthetic gene circuit based on a heterologous pair consisting of a promoter and a transcription factor known to respond to the metabolite. We exploited the met operator and MetJ repressor of Escherichia coli, the interaction between which depends on S-adenosylmethionine (SAM), to construct synthetic gene circuits that report SAM levels in Saccharomyces cerevisiae. Using a dual-input circuit that outputs selection marker genes in a doxycycline-tunable manner, we screened a genomic library to identify GAL11 as a novel multicopy enhancer of SAM levels. These results demonstrate the potential and utility of synthetic gene circuit-mediated metabolite monitoring.",
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AU - Ito, Takashi

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