Fig. 2: Rational design and evaluation of complex multicellular patterns through specific synthetic adhesion in yeast.
From: Programmable cell–cell adhesion in synthetic yeast communities for improved bioproduction
a–c, Schematic representation and confocal microscopy images demonstrating adhesion specificity using a nonadhesion reference cell 3 (blue). Figures present representative fluorescent images from experiments that were independently repeated at least three times with similar outcomes. a, In the control strain with seven adhesion genes deleted (FLO1, FLO5, FLO10, FLO11, AGA1, AGA2, SAG1), all cell types remain dispersed. Microscopy images (strains): 007Δ sfGFP (green cells), 007Δ mRuby2 (red cells), 007Δ mTagBFP2 (blue cells). b, Expression of FLO1 or FLO5 in cell 1 and cell 2 results in nonspecific adhesion to cell 3. Microscopy images (strains): 007Δ FLO1 sfGFP (green cells), 007Δ FLO1 mRuby2 (red cells), 007Δ mTagBFP2 (blue cells). c, Specific adhesion pairs (Strategy 2) mediate adhesion between cell 1 and cell 2, but not to cell 3. Microscopy images (strains): 007Δ SAG1 sfGFP (green cells), 007Δ AGA1 AGA2 mRuby2 (red cells), 007Δ mTagBFP2 (blue cells). Strategy 2.1 was selected as an example of Strategy 2. Strategy 2.2 showed a similar pattern and can be found in Supplementary Fig. 3e. d, Quantification of the percentage of each cell cluster type after introducing cell 3 (blue) across various adhesion systems (Strategy 1, Strategy 2.1 and Strategy 2.2). Tricolor cell clusters are cell clusters containing all three fluorescent colors (green, red and blue); bicolor clusters with blue are cell clusters containing two colors, one of which is blue; cluster with red and green cells are cell clusters containing only red and green cells; and monochrome cells are single-colored cells. Quantification was determined by calculating the ratio of each cell type to the total cell count, derived from six confocal images obtained from confocal sections of n = 3 biologically independent replicates. e, Typhoon biomolecular imager analysis of 2D pattern formation using engineered yeast cells expressing FLO1. Cell 2 (007Δ FLO1 mRuby2) adheres to cell 1 (007Δ FLO1 sfGFP), replicating the pattern of cell 1. Scale bars, 50 µm.
