Fig. 3: Design of ZFDs for the loxF8 genomic locus.
From: Activation of recombinases at specific DNA loci by zinc-finger domain insertions
a, Genomic sequences flanking loxF8. Binding sites of the designed ZFDs upstream (orange, ZFL1–2) and downstream (blue, ZFR1–4) are shown with the distance (bp) between the loxF8 and the ZF motifs indicated. b, Representative plasmid-based activity assay of the monomers from the D7 recombinase heterodimer (D7L and D7R) fused with the designed ZFDs on the symmetric loxF8L and loxF8R target sites and their extended versions that include the respective flanking genomic sequences for ZFD binding (loxF8L-flank and loxF8R-flank). The activity of the wild-type monomers is shown as a control. High induction levels (100 µg ml−1 ʟ-arabinose for D7L and D7L–ZFL; 200 µg ml−1 ʟ-arabinose for D7R and D7R–ZFR) were used. The upper band represents the unrecombined plasmid (line with two triangles), and the lower band represents the recombined plasmid (line with one triangle). The assay was performed three times (n = 3, biologically independent samples; replicates are shown in Source Data files). M, marker. Fusions with activity (D7L–ZFL1 and D7R–ZFR4) are highlighted with an orange box and a blue box, respectively.
