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. 2016 Mar 7;8(3):72. doi: 10.3390/v8030072

Figure 2.

Figure 2

Figure 2

gRNA guided Cas9 induces homologous recombination by creating a DNA double-stranded break in the target region of vaccinia virus. (A) The gRNA is transcribed; (B) gRNA guides Cas9 to the target site; (C) Cas9 creates a DNA double-stranded break, which is repaired by the repair donor vector via the mechanism of homologous recombination; and (D) the mutant vaccinia virus is generated with the deletion of the target gene and insertion of the purification marker RFP containing its promoter H5. A therapeutic gene hGM-CSF and its promoter H5 can also be incorporated into the target gene.