Extended Data Fig. 1: Development and optimization of the CaTCH reporter.

a, Functional comparison of dCas9-VPR to dCas9-VP64 using a doxycycline-inducible reporter system with sgRNAs targeting either one (−53 bp or −203 distance to TSS), two, or seven target sites (tetO sites). Doxycycline induced activation via rtTA3 was used as a positive control. Reporter activation was measured by FACS. Left: Corresponding FACS histograms of GFP activation (normalized to mode); Right: Quantification of percent activated of parent (upper panel) and of signal strength in mean fluorescent intensity (MFI, lower panel), n = 3 biologically independent samples, for CTRL n = 2, bar graphs display the mean ± standard deviation (s.d.). b, Evaluating the optimal sgRNA positioning using two reporter constructs (D1 and D2). Left: FACS histograms of GFP activation with the individual sgRNAs in D1 or D2, ordered by sgRNA-distance to the TSS (−58 bp, −82 bp, −106 bp, −130 bp, −154 bp, −202 bp, −250 bp, −298 bp); Right: Quantification of percent activated and MFI; n = 2 biologically independent samples, bar graphs display the mean. c, Determining the effect of multiple sgRNA target sites and their spacing on GFP activation by comparing a reporter construct with spaced BCs (R1) with a reporter construct with BCs side by side (R2). Left: FACS histograms of GFP activation; Right: Quantification of percent activated and MFI, n = 2 biologically independent samples, bar graphs display the mean.