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HMEC-1 endothelial cell line expressing a synthetic gene network which produces enhanced green fluorescent protein (eGFP) in response to wall shear stress applied on the cultured cell monolayer.

 

 

The mechanosensitive synthetic gene network consists of a sensor module, a reporter module, and a linker module that signals between the sensor and reporter. The sensor module is composed of the Bradykinin B2 G-PCR linked to the tetracycline-controlled transcriptional trans-activator (tTA).The linker between the GPCR and tTA is a 7 amino acid long Tobacco etch virus (TEV) protease cleavage site. The reporter module contains eGFP under the control of a tTA dependent promoter. The linker module is βArrestin2 signalling protein fused to the TEV protease enzyme. Upon activation of the G-PCR by shear stress or by an extracellular ligand (bradykinin), βArrestin2 binds to the cytoplasmic domain of the G-PCR, causing the cleavage of the linker between tTA and the G-PCR by the TEV protease. Consequently, the released tTA promotes transcription from the reporter module.

Internal case number: 7553

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Mechanosensing Fluorescent Cell Line

Reporter cell line in which GFP expression is stimulated in response to mechanical force.

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