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. 2014 Mar 10;136(13):5047–5056. doi: 10.1021/ja4130059

Figure 4.

Figure 4

Site-specific integration into the human genome by enhanced ZFRs. (a) Schematic representation of sequence and location of the ZFR target site on human chromosome 4. Black triangles indicate cleavage sites within DNA target. Red line denotes the approximate position of the ZFR target site. (b) Bulk PCR analysis of HEK293 cells transfected with an empty donor plasmid containing only a puromycin-resistance gene and various ZFR pairs designed to target human chromosome 4. Integration was evaluated in the forward and reverse orientations. GAPDH indicates PCR control. DO indicates donor only (no eZFRs). Genome-wide integration rates indicated beneath each lane. (c) Clonal PCR analysis of puromycin-resistant cells transfected with empty donor and eZFRs in both orientations.