Hypoxia-responsive transgene expression system using RTP801 promoter and synthetic transactivator fused with oxygen-dependent degradation domain

Akihiko Ono, Akira Ito, Tomonaga Sato, Masaki Yamaguchi, Taiga Suzuki, Kawabe Yoshinori, Masamichi Kamihira

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Precise control of gene expression using an artificial gene circuit is a major challenge in the application of synthetic biology. Here, we designed a hypoxia-responsive transgene expression system by combining a hypoxia-inducible RTP801 promoter and a tetracycline-responsive transactivator fused with an oxygen-dependent degradation domain (TA-ODD). The reporter gene expression was highly induced by hypoxia when a transactivator-expression plasmid, pRTP801/TA-ODD, harboring a TA-ODD gene driven by the RTP801 promoter, was cotransfected with a reporter plasmid, pTRE/EGFP, harboring an EGFP gene controlled under the transactivator-responsive promoter. A stable cell line into which the expression units RTP801/TA-ODD and TRE/EGFP had been introduced responded to hypoxia and expressed the reporter gene in an oxygen-concentration-dependent manner. Moreover, the cells demonstrated potential as sensors to detect hypoxic conditions in a three-dimensional tissue culture in vitro. These results indicate that the hypoxia-responsive transgene expression system is useful for constructing cell-based hypoxia detection systems.

Original languageEnglish
Pages (from-to)115-124
Number of pages10
JournalJournal of Bioscience and Bioengineering
Volume124
Issue number1
DOIs
Publication statusPublished - Jul 1 2017

Fingerprint

Trans-Activators
Transgenes
Oxygen
Degradation
Genes
Gene expression
Reporter Genes
Plasmids
Tissue culture
Synthetic Biology
Synthetic Genes
Gene Expression
Cell Hypoxia
Gene Regulatory Networks
Tetracycline
Cells
Hypoxia
Networks (circuits)
Sensors
Cell Line

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Cite this

Hypoxia-responsive transgene expression system using RTP801 promoter and synthetic transactivator fused with oxygen-dependent degradation domain. / Ono, Akihiko; Ito, Akira; Sato, Tomonaga; Yamaguchi, Masaki; Suzuki, Taiga; Yoshinori, Kawabe; Kamihira, Masamichi.

In: Journal of Bioscience and Bioengineering, Vol. 124, No. 1, 01.07.2017, p. 115-124.

Research output: Contribution to journalArticle

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AU - Yoshinori, Kawabe

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