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. 2014 May 14;9(5):e97579. doi: 10.1371/journal.pone.0097579

Figure 2. HBV-ZFN activity in mammalian cells.

Figure 2

(a) HBV-ZFN target site/GFP reporter plasmid containing a ZFN target site (red) placed immediately after the GFP start codon (blue) and preceding the in-frame GFP ORF (green). The HBV-ZFN spacer is shown in lower case. (b) ZFN reporter constructs and expression plasmids for ZFN pairs 1–3 were transfected into HEK 293T cells. GFP expression was visualized at 72 hours post transfection. (c) ZFN-mediated target site disruptions within reporter constructs were amplified by PCR from the HEK 293T cells that had been transfected with ZFN-expressing plasmids as indicated. Indels found at each ZFN site are aligned with the wild type target sequence (wt) and nucleotides with differences in at least one sequence are shown in color. The spacer regions are underlined. GFP – green fluorescent protein; SFFV – spleen focus-forming virus promoter; WPRE – woodchuck hepatitis post-transcriptional regulatory element; polyA – polyadenylation signal; wt – wild type; ZFN – zinc finger nuclease.