DNA damage-responsive transgene expression mediated by the p53 promoter with transcriptional amplification

doi:10.1016/j.jbiosc.2015.02.009

Journal of Bioscience and Bioengineering

Volume 120, Issue 4, October 2015, Pages 463-466

https://doi.org/10.1016/j.jbiosc.2015.02.009 Get rights and content

We constructed a DNA damage-responsive transgene expression system mediated by the p53 promoter. We incorporated a transactivation system to generate transcriptional amplification via a positive feedback loop. Higher levels of DNA damage-responsive transgene expression were observed when transactivation was active.

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Cited by (5)

Hypoxia-responsive expression of vascular endothelial growth factor for induction of angiogenesis in artificial three-dimensional tissues
2021, Journal of Bioscience and Bioengineering
Citation Excerpt :
Additionally, abundant ring-like structures were observed in images of vascular endothelial cell staining of tissues that consisted of C2C12/hypoVEGF[#5] cells (Fig. 5B). Previously, we had developed gene expression systems to detect cellular environments, such as heat (16,17), cell damage (18), and hypoxia (7), by a synthetic biological approach (19). In this study, we designed a hypoxia-responsive VEGF gene expression system.
Constructing three-dimensional (3D) tissues is an important process to improve cellular functions in tissue engineering. When transplanting artificially constructed tissues, a poor vascular network restricts oxygen and nutrient supplies to the tissue cells, which leads to cell death and reduced rates of tissue engraftment. Therefore, it is necessary to develop a system that builds a vascular network within 3D tissues. Here, we developed a hypoxia-responsive gene expression system for production of an angiogenic factor, vascular endothelial growth factor (VEGF), to improve hypoxia and nutrition deficiencies inside artificial 3D tissues. We demonstrated that cells into which the hypoxia-responsive VEGF gene expression system had been introduced autonomously controlled VEGF expression in a hypoxic stress-dependent manner. Next, we confirmed that VEGF expression within a 3D cell sheet was induced in response to a hypoxic environment in vitro. The genetically modified cell sheet was subcutaneously transplanted into mice to evaluate the feasibility of the hypoxia-responsive VEGF gene expression system in vivo. The results suggest that the hypoxia-responsive VEGF gene expression system is promising to prepare artificial 3D tissues in regenerative medicine.
Hypoxia-responsive transgene expression system using RTP801 promoter and synthetic transactivator fused with oxygen-dependent degradation domain
2017, Journal of Bioscience and Bioengineering
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.
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References (17)

Heat-inducible transgene expression system incorporating a positive feedback loop of transcriptional amplification for hyperthermia-induced gene therapy

J. Biosci. Bioeng.

A genotoxicity test system based on p53R2 gene expression in human cells: assessment of its reactivity to various classes of genotoxic chemicals

Mutat. Res.

Induction of chromatid breakage by clastogens in cells in G2 phase

Mutat. Res.

Genotoxicity test system based on p53R2 gene expression in human cells: examination with 80 chemicals

Mutat. Res.

Oncogenic mutations of the p53 tumor suppressor: the demons of the guardian of the genome

Cancer Res.

Live or let die: the cell's response to p53

Nat. Rev. Cancer

Induction of nuclear accumulation of the tumor-suppressor protein p53 by DNA-damaging agents

Oncogene

Identification of a novel p53 promoter element involved in genotoxic stress-inducible p53 gene expression

Mol. Cell Biol.

Cited by (5)

Hypoxia-responsive expression of vascular endothelial growth factor for induction of angiogenesis in artificial three-dimensional tissues

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

RNA Aptamer-Mediated Gene Activation Systems for Inducible Transgene Expression in Animal Cells

Cell Engineering Technology for Creating Functional Animal Cells

Magnetically triggered transgene expression system using magnetic nanoparticles

Technical NoteDNA damage-responsive transgene expression mediated by the p53 promoter with transcriptional amplification

J. Biosci. Bioeng.

Mutat. Res.

Mutat. Res.

Mutat. Res.

Oncogenic mutations of the p53 tumor suppressor: the demons of the guardian of the genome

Cancer Res.

Live or let die: the cell's response to p53

Nat. Rev. Cancer

Induction of nuclear accumulation of the tumor-suppressor protein p53 by DNA-damaging agents

Oncogene

Identification of a novel p53 promoter element involved in genotoxic stress-inducible p53 gene expression

Mol. Cell Biol.

Technical Note
DNA damage-responsive transgene expression mediated by the p53 promoter with transcriptional amplification