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Spatiotemporal control of integrin-mediated cell adhesions to extracellular matrix regulates cell behavior with has numerous implications for biotechnological applications. In this work, two approaches for regulating cell adhesions in space and time with high precision are reported, both of which utilize green light. In the first design, CarH, which is a tetramer in the dark, is used to mask cRGD adhesion-peptides on a surface. Upon green light illumination, the CarH tetramer dissociates into its monomers, revealing the adhesion peptide so that cells can adhere. In the second design, the RGD motif is incorporated into the CarH protein tetramer such that cells can adhere to surfaces functionalized with this protein. The cell adhesions can be disrupted with green light, due to the disassembly of the CarH-RGD protein. Both designs allow for photoregulation with noninvasive visible light and open new possibilities to investigate the dynamical regulation of cell adhesions in cell biology.

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Cell adhesions to the extracellular matrix and to neighboring cells are fundamental to cell behavior and have also been implemented into minimal synthetic cells, which are assembled from molecular building blocks from the bottom-up. Investigating adhesion in cell mimetic models with reduced complexity provides a better understanding of biochemical and biophysical concepts underlying the cell adhesion machinery. In return, implementing cell-matrix and cell-cell adhesions into minimal synthetic cells allows reconstructing cell functions associated with cell adhesions including cell motility, multicellular prototissues, fusion of vesicles, and the self-sorting of different cell types. Cell adhesions have been mimicked using both the native cell receptors and reductionist mimetics providing a variety of specific, reversible, dynamic, and spatiotemporally controlled interactions. This review gives an overview of different minimal adhesion modules integrated into different minimal synthetic cells drawing inspiration from cell and colloidal science.

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The dynamic and spatiotemporal control of integrin-mediated cell adhesion to RGD motifs in its extracellular matrix (ECM) is important for understating cell biology and biomedical applications because cell adhesion fundamentally regulates cellular behavior. Herein, the first photoswitchable synthetic ECM protein, Photo-ECM, based on the blue light switchable protein LOV2 is engineered. The Photo-ECM protein includes a RGD sequence, which is hidden in the folded LOV2 protein structure in the dark and is exposed under blue light so that integrins can bind and cells can adhere. The switchable presentation of the RGD motif allows to reversibly mediate and modulate integrin-based cell adhesions using noninvasive blue light. With this protein cell adhesions in live cells could be reversed and the dynamics at the cellular level is observed. Hence, the Photo-ECM opens a new possibility to investigate the spatiotemporal regulation of cell adhesions in cell biology and is the first step toward a genetically encoded and light-responsive ECM.

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Biofilms are not only a leading cause of chronic infections and biofouling, but they also have a tremendous positive potential in biotechnology for biocatalysis and waste treatment. Biofilms are spatially structured communities of microbes, which exchange chemicals and communicate with each other. By spatially controlling bacterial adhesion to surfaces, and therefore the microstructure of biofilms, a promising method of understanding social interactions between bacteria and designed biofilms is developed. The bacterial photolithography approach described here allows to photopattern specific bacteria adhesion molecules, to control surface adhesion, and to guide the formation of biofilms. To do this, α-D-mannoside, which is recognized by the Escherichia coli FimH receptor, is linked to a nonadhesive polyethylene glycol surface through a photocleavable 2-nitrobenzyl linker. When a pattern of UV light in a specific shape is projected onto these surfaces, the light-exposed areas become nonadhesive and bacteria only adhere to the dark, unexposed areas in the photopattern. Bacterial photolithography enables bacterial patterning with high spatial resolution down to 10 µm without mechanical interference. Additionally, patterning biofilms with complicated geometries allows studying the importance of microscale spatial organization on the collective behavior of bacteria such as quorum sensing.

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We use plasmon rulers to follow the conformational dynamics of a single protein for up to 24 h at a video rate. The plasmon ruler consists of two gold nanospheres connected by a single protein linker. In our experiment, we follow the dynamics of the molecular chaperone heat shock protein 90 (Hsp90), which is known to show "open" and "closed" conformations. Our measurements confirm the previously known conformational dynamics with transition times in the second to minute time scale and reveals new dynamics on the time scale of minutes to hours. Plasmon rulers thus extend the observation bandwidth 3-4 orders of magnitude with respect to single-molecule fluorescence resonance energy transfer and enable the study of molecular dynamics with unprecedented precision.

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Independent control over multiple cell-material interactions with high spatiotemporal resolution is a key for many biomedical applications and understanding cell biology, as different cell types can perform different tasks in a multicellular context. In this study, the binding of two different cell types to materials is orthogonally controlled with blue and red light providing independent regulation in space and time. Cells expressing the photoswitchable protein cryptochrome 2 (CRY2) on cell surface bind to N-truncated CRY-interacting basic helix-loop-helix protein 1 (CIBN)-immobilized substrates under blue light and cells expressing the photoswitchable protein phytochrome B (PhyB ) on cell surface bind to phytochrome interaction factor 6 (PIF6)-immobilized substrates under red light, respectively. These light-switchable cell interactions provide orthogonal and noninvasive control using two wavelengths of visible light. Moreover, both cell-material interactions are dynamically switched on under light and reversible in the dark. The specificity of the CRY2/CIBN and PhyB/PIF6 interactions and their response to different wavelengths of light allow selectively activating the binding of one cell type with blue and the other cell type with red light in the presence of the other cell type.