Background-suppressed live visualization of genomic loci with an improved CRISPR system based on a split fluorophore

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Figure 2.
Figure 2.

SunTag split-sfGFP CRISPR system reveals target-specific foci with high S/B. (A) C9-1 region observed in a fixed stable AD-293 cell line expressing the SunTag split-sfGFP CRISPR system. Schematic shows the location of the C9-1 loci on Chromosome 9. (B) Simultaneous CRISPR and DNA FISH imaging of the C9-1 loci to verify the colocalization of GFP foci and DNA FISH foci. (C) Comparison of signal, background, and S/B in C9-1 region observed with SunTag sfGFP, MCP-mCherry, and SunTag split-sfGFP systems. BG stands for background. Schematic shows how S/B was defined from the images. (D) Distribution of fluorescence signal intensity of the detected C9-1 foci in fixed stable AD-293 cell lines expressing the SunTag sfGFP or SunTag split-sfGFP systems. (E) Histograms of the number of C9-1 foci detected with different labeling schemes.

This Article

  1. Published in Advance September 4, 2020, doi: 10.1101/gr.260018.119 Genome Res. 30: 1306-1316

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