Extended Data Fig. 1: Light-inducible gene activation and knock-down modules.
From: Spatiotemporal, optogenetic control of gene expression in organoids
a. Log2 fold-change of the ASCL1 mRNA, induced with the CPTS (left) and SCPTS (right) systems3, between a non-targeting (NT) and promoter-targeting sgRNA (ASCL1), in dark or lit conditions at 24h, measured by RT-qPCR. Each dot represents a biological replicate (n = 2). b. Histograms of pixel intensity with and without background correction (green and red), performed with the subtract background function in Fiji/imageJ with a rolling ball radius of 50, for three representative samples of the SCPTS CaSP2 CasRx system (left: 6h time point with non-targeting guide, middle: 50h time point with the Tet6 targeting guide in the dark, right: 50h time point with the Tet6 targeting guide with illumination). c. Background-subtracted mean GFP intensity, in either dark or lit conditions (n = 4) for constitutive CasRx. P-values (Benjamini-Hochberg corrected two-sided t test) between dark and lit conditions at 50h are shown. d-e. As in Fig. 1d, e, for an additional time point at 96h (n = 3, except for NT guide control n = 1). Each dot represents a replicate (n = 3, except for NT guide n = 1) and horizontal lines represent means over each condition. P-values (Benjamini-Hochberg corrected two-sided t test) between dark and lit condition for CasRx guide and dox/no dox conditions are shown. f. Brighfield images corresponding to the panels in Fig. 1f, g. Scale bar (top left panel, applies to all): 50 μm. Horizontal lines represent the mean of all replicates in each plot.
