Figure 4: Restoration of FnCas9 nuclease activity in K562 cells by catalytically dead SpCas9 (SpdCas9) binding at proximal locations. | Nature Communications

Figure 4: Restoration of FnCas9 nuclease activity in K562 cells by catalytically dead SpCas9 (SpdCas9) binding at proximal locations.

From: Targeted activation of diverse CRISPR-Cas systems for mammalian genome editing via proximal CRISPR targeting

Figure 4

(a) Schematic of a proximity chromatin interference hypothesis. FnCas9 is unable to access an endogenous target in a certain chromatin configuration, but the binding of SpdCas9 at proximal locations alters the local chromatin homeostasis and enables FnCas9 to access and cleave the otherwise inaccessible target. (b) FnCas9 and SpdCas9 targets in the POR exon 8 genomic region for testing the hypothesis. Targets are indicated by bars and PAMs are highlighted in purple. FnCas9 cleavage positions are indicated by yellow triangles. (c) FnCas9 target DNA cleavage activities under different conditions. The binding of SpdCas9 at one proximal site restored FnCas9 cleavage activity and two SpdCas9 proximal binding sites acted synergistically. Data are representative of three independent experiments. The sgRNA numbers correspond to the target numbers in b. M, wide-range DNA markers; ND, not determined.

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