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
frst 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 fow
setup that exposes cells to linearly increasing shear stress along the length of the fow channel foor.
Activation of the gene network varied logarithmically as a function of shear stress magnitude.