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. 2017 Jul 21;7:6122. doi: 10.1038/s41598-017-05905-w

Figure 6.

Figure 6

The effect of nickase-Cas9 with the pair of sgRNAs in vivo. (A) Schematic of the split LUxxUC plasmid inserted into the target HBc sequence and pU6-sgRNA-HBc-1/2 targeting the HBc sequence. A recombination of firefly luciferase is induced by DSBs on the inserted targeted sequence. (B) Cas9, nicakse-Cas9 and d-Cas9 with a pair of sgRNA-HBc-1/2-expressing plasmids were transfected with split LUxxUC plasmid into 293T cells. The firefly luciferase activity was determined at 24 hr post -transduction. (C,D) Cas9 and nickase-Cas9 plus the pair of sgRNA-HBc-1/2 or d-Cas9 plus the pair of sgRNA-HBc-1/2 with split LUxxUC-expressing plasmids were delivered into the livers of C57BL/6j mice via hydrodynamic injection (n = 3 mice per group). Luciferase activity was detected at 2 days post injection. The luciferase intensity was measured by an IVIS system. *p < 0.01 vs. the results for control cells.