Fig. 2: Validation of Casilio imaging specificity. | Nature Communications

Fig. 2: Validation of Casilio imaging specificity.

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

Fig. 2

a Schematic showing locations of Casilio MUC4 gRNA (gMUC4) and Cy5-MUC4 FISH probes. Chr, chromosome. b Co-labeling of MUC4 locus by Casilio (Green) and DNA-FISH (Cy5-MUC4, Red) in U2OS cells (n = 8 nuclei; 28 spots; Specificity = #Red-Green-overlap/#Green=96%). Scale bar, 5 μm. c Co-labeling of Casilio (Green) and DNA-FISH (Cy5-MUC4, Red) in ARPE-19 cells (n = 16 nuclei; 40 spots; Specificity = 100%). Scale bar, 5 μm. d Percentage of green ARPE-19 nuclei with the indicated numbers of Casilio/gMUC4 spots in live cells without and with dCas9. Nucleus counts are indicated on top of the columns. e Percentage of green HAP1 nuclei with the indicated numbers of Casilio/gMUC4 spots in live cells. Nucleus counts are indicated on top of the columns. Source data are provided as a Source Data file.

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