Development of a synthetic gene network to modulate gene expression by mechanical forces

Authors

Zoltan Kis, Imperial College London
Tania Rodin, Imperial College London
Asma Zafar, Imperial College London
Zhangxing Lai, Imperial College London
Grace Freke, Imperial College London
Oliver Fleck, Imperial College London
Armando Del Rio Hernandez, Imperial College London
Leila Towhidi, Imperial College London
Ryan M. Pedrigi, University of Nebraska-LincolnFollow
Takayuki Homma, Imperial College London
Rob Krams, Imperial College LondonFollow

Date of this Version

2016

Citation

Scientific Reports | 6:29643

Comments

Open access

doi: 10.1038/srep29643

Abstract

The majority of (mammalian) cells in our body are sensitive to mechanical forces, but little work has been done to develop assays to monitor mechanosensor activity. Furthermore, it is currently impossible to use mechanosensor activity to drive gene expression. To address these needs, we developed the first mammalian mechanosensitive synthetic gene network to monitor endothelial cell shear stress levels and directly modulate expression of an atheroprotective transcription factor by shear stress. The technique is highly modular, easily scalable and allows graded control of gene expression by mechanical stimuli in hard-to-transfect mammalian cells. We call this new approach mechanosyngenetics. To insert the gene network into a high proportion of cells, a hybrid transfection procedure was developed that involves electroporation, plasmids replication in mammalian cells, mammalian antibiotic selection, a second electroporation and gene network activation. This procedure takes 1 week and yielded over 60% of cells with a functional gene network. To test gene network functionality, we developed a flow setup that exposes cells to linearly increasing shear stress along the length of the flow channel floor. Activation of the gene network varied logarithmically as a function of shear stress magnitude.