While ultrasound was used to stimulate mechanosensors in our project, since it is most appropriate for the stimulation of deep tissue, other mechanical stimuli could also trigger activation of mechanosensors. Mechanical stimulation appears to be a critical modulator for many aspects of biology, both of living tissue and cells Seriani2016. Mechanosensing can cell proliferation and differentiation Shah2014. In recent years, many methods of activation of mechanosensitive channels have been implemented. Among the activators of mechanosensors, hyperosmolarity has been shown to trigger the activation of certain genes Veltmann. Additional mechanical stimuli such as fluid shear stress causes activation of embryonic endothelial cells Ranade2014.

Therefore our system might be also activated by other mechanical stimuli. One of them was stimulation by direct contact that underlies the sense of touch. Our designed mechano-responsive system is composed of two modules. First we included modules to provide and increase the sensitivity to mechanical stress by mechanoresponsive ion channels and gas vesicles and secondly, the influx of calcium is visualized by the dimerization of calmodulin (attached to the C terminus of split luciferase) and M13 (attached to the N terminus of split luciferase) which results in bioluminescence after the reconstitution of split luciferase. This means that this system is able to convert the mechanical stimulus into light signal, or to put it differently, with our system we are able to see when cells are touched.

Shear flow also exerts mechanical force on cells, which also has relevance for the endothelial cells within blood vessels, therefore also the flow of the liquid medium around cells might be sensed by cells. The experiment to test this ability was performed by agitating the petri dish with attached cells. In order to identify if the engineered cells are able to respond to the sheaf flow we included control cells that either constitutively express the luciferase and cells that harbour only the calcium-dependent reporter without constructs to increase the mechanosensing along with cells expressing the gas vesicles and reporters.

Results show that the light was emitted only in cells with the constitutive luciferase if the plate has not been moved(1>/ref>). On the other hand shaking triggered activation of the luciferase, with significantly higher level in cells that harboured gas vesicles in addition to the reporter, demonstrating that activation does not require high pressure on cells.