RRC ID 52129
Author Ono A, Ito A, Sato T, Yamaguchi M, Suzuki T, Kawabe Y, Kamihira M.
Title Hypoxia-responsive transgene expression system using RTP801 promoter and synthetic transactivator fused with oxygen-dependent degradation domain.
Journal J Biosci Bioeng
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.
Volume 124(1)
Pages 115-124
Published 2017-7-1
DOI 10.1016/j.jbiosc.2017.02.012
PII S1389-1723(16)30572-2
PMID 28284789
MeSH Amino Acid Sequence Animals Base Sequence Cell Hypoxia / genetics Gene Expression Genes, Reporter / genetics Genetic Engineering / methods* HeLa Cells Humans Mice Oxygen / metabolism* Plasmids / genetics Promoter Regions, Genetic / genetics* Protein Domains Trans-Activators / genetics* Transcription Factors / chemistry* Transcription Factors / genetics* Transcription Factors / metabolism Transgenes / genetics
IF 2.366
Times Cited 2
Human and Animal Cells C2C12(RCB0987) HeLa