Fig. 4: CRISPR-SunTag activators form phase-separated condensates.
From: Dynamic properties of transcriptional condensates modulate CRISPRa-mediated gene activation
a Schematic illustrating the fusion of GFP to dCas9 for imaging its subcellular localization in different CRISPRa systems. b Representative images depicting dCas9-GFP puncta formed in each CRISPRa system before or after 1.5% 1,6-hexanediol (1,6-HD) for 2 min. Scale bars, 5 μm. GFP punta number and total signal intensity of puncta per cell were quantified for individual conditions. VPR: n = 41, VPRF: n = 48, 3xVPR: n = 50, 10xVPR: n = 54, 3xVPRF: n = 48, 10xVPRF: n = 42, PCP-12xVPRF: n = 52, PCP-40xVPRF: n = 50. All plots represent mean ± SEM; P values were calculated by unpaired two-tailed t-test. c Representative images of the FRAP experiment. Bleaching was performed at the indicated time points. A representative puncta was indicated with a white box. Images were recorded with 10 s intervals. Scale bars, 5 μm. d FRAP recovery curves showing the GFP intensity overtime in the bleach region in (c). The mobile fraction (Fm) and recovery half time (1/2 t) were calculated for the recovery curves. Data are shown as mean ± SEM. The number of cells quantified in (d), from left to right, is 10, 20, 12, 12, 10, 15, 12 and 13, respectively. Source data are provided as a Source Data file.
