Fig. 5: Three-color three-point live-cell imaging of chromatin loops. | Nature Communications

Fig. 5: Three-color three-point live-cell imaging of chromatin loops.

From: CRISPR-mediated multiplexed live cell imaging of nonrepetitive genomic loci with one guide RNA per locus

Fig. 5

a Casilio probes for visualizing chromatin interactions of IER5L promoter and its super-enhancer, with locus P (promoter) labeled by gRNA-20xPBS48107/Clover-PUF48107, locus M (mid-point) labeled by gRNA-20xPBSc/iRFP670-PUFc, and locus SE (super-enhancer) labeled by gRNA-25xPBS9R/PUF9R-mRuby2. Chr, chromosome. b, c Representative nuclei (nuclear boundaries outlined) and time-lapse images of IER5L-P-SE loop, with IER5L promoter, midpoint and super-enhancer labeled by Clover (green, stealth arrowhead), iRFP670 (red, triangle arrowhead), mRuby2 (magenta, open arrowhead), respectively, in HCT116/RAD21-mAID cells at the indicated time point. Scale bars 5 μm. See Supplementary Videos 6, 7. See Supplementary Fig. 6 for distance plots of imaged nuclei (n = 18 nuclei; 28 alleles). Experiment repeated 7 times.

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