Publication date: 25th July 2016
Cells interact with the extracellular matrix and neighboring cells forming cell-cell and cell-matrix contacts, which are mediated by cadherins and integrins, respectively and the expression of them is both spatially and temporally tightly regulated during many biological processes including embryogenesis and EMT-MET. Our aim is to generate platforms where we can control cell-cell and cell-surface contacts spatially, temporally and reversibly using visible light responsive proteins from plants (PhyB-PIF6 and CRY2-CIBN heterodimers) to provide a better understanding of cell contacts. To mimic cell-cell contacts, the light dependent heterodimers, CRY2-CIBN and PhyB-PIF6 are employed. CRY2 is switched to its photoactive state with blue light, which allows CRY2 to bind CIBN. This binding is reversed passively in dark. Similarly, red light illumination activates PhyB to bind PIF6 and this interaction is reversed under far-red light or in dark. We expressed complementary heterodimers on the surfaces of cells, which do not express cadherins, and can trigger and break the cell-cell contacts in light/dark cycles. Towards controlling cell-surface contacts, we used PEG functionalized glass surfaces where we immobilized one of the heterodimerization partners (CIBN or PIF6). By incubating the cells expressing the complementary heterodimer (CRY2 or PhyB) in dark and red/blue light illumination on the surface, we can turn on or off cell-surface contacts. These platforms give us high spatial and temporal control over cell-matrix and cell-cell contacts in a non-invasive fashion and will provide new insight into the interplay of these cell contacts in biological processes.
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