| 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
|
| Resource |
| Human and Animal Cells |
C2C12(RCB0987)
HeLa |
