Extended Data Fig. 1: Generation of blue–light-inducible AKP colon organoids.

a, Schematic of the plasmids comprising the OptoCre module. The promoter, gene, and restriction sites used to generate the plasmids are indicated. b, Schematic of the integration of the different genetic elements that allow spatiotemporal control of oncogenic recombination in colon organoids. c, Schematic of the experimental workflow used to test and optimize the OptoCre system. d, Brightfield and fluorescence images of OptoCre-carrying inducible colon organoids exposed to the indicated conditions. Red and green signals correspond to healthy and mutated cells, respectively. Images were taken 48 h after induction. Scale bar, 200 μm. e, Recombination efficiency in inducible colon organoid exposed to the conditions indicated in panel d. ***P = 0.0001 (Kruskal-Wallis and Dunn’s multiple comparisons test; n = 9 cultures for each condition). Each point represents one well of organoids. Data represent mean ± SEM. f, Brightfield and fluorescence images of inducible colon organoids exposed to control (top) and activation (bottom) conditions, dissociated into single cells and replated in the absence of growth factors (BMGF medium). Green signal corresponds to mutated cells. Images were taken 24 h after replating. Scale bar, 200 μm.

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