Fig. 3: Parallel orthogonal circuits for quantitative cell-state control.
a, Design of a 2× orthogonal branching circuit using CA and GA att sites, distinct promoters and fluorescent reporters. b, Theoretical progeny states and probabilities: DD (ShadowG only), GD (Venus), RG (Venus + mScarlet) and RD (mScarlet). c, Agreement between calculated and measured progeny distributions by flow cytometry (mean ± s.d., n = 8 biological replicates). d, Fluorescence images showing four cell states in liquid culture and colonies; representative images from three independent experiments. Scale bars, 10 μm (left); 1 mm (right). e, Design of a 3× orthogonal branching circuit. f, Theoretical progeny states and probabilities. g, Comparison of predicted and observed distributions by flow cytometry (mean ± s.d., n = 8 biological replicates). h, Fluorescence images showing eight cell states in liquid culture and colonies; representative images from three independent experiments. Scale bars, 10 μm (left); 1 mm (right). i–k, Schematic designs, predicted distributions and flow-cytometry data for 1× (i), 2× (j) and 3× (k) parallel branching circuits (mean ± s.d., n = 8 biological replicates).
