Extended Data Fig. 3: Activity tests of ZF-dependent designer-recombinases.
From: Activation of recombinases at specific DNA loci by zinc-finger domain insertions
a Plasmid-based activity assay for Brec1278-ZFCCR5L, in which the ZifCCR5L is fused between the residues 278 and 279 of Brec1 via the (GGS)8 linkers. The fusion complex was tested on loxBTR, loxBTR-5-zifCCR5L (A), or loxBTR-5-zifCCR5L (B) target sites. The test was performed at 200 ug/ml L-arabinose. Activity of the wild-type Brec1 is shown as a control. b Plasmid-based activity assay of RecHTLV278-Zif268 fusion complex, in which the Zif268 is fused between the residues 278 and 279 of RecHTLV via the (GGS)8 linkers. The fusion complex was tested on loxHTLV, loxHTLV-5-zif268 (A), or loxHTLV-5-zif268 (B) target sites. The test was performed at 100 ug/ml L-arabinose. Activity of the wild-type RecHTLV is shown as a control. c Plasmid-based activity assay for D7L278-Zif268, in which the Zif268 is fused between the residues 278 and 279 of D7L via the (GGS)8 linkers. The fusion complex was tested on loxF8L, loxF8L-5-zif268 (A), or loxF8L-5-zif268 (B) target sites. The test was performed at 10 ug/ml L-arabinose. Activity of the wild-type D7L is shown as a control. d Plasmid-based activity assay for D7R278-Zif268, in which the Zif268 is fused between the residues 278 and 279 of D7R via the (GGS)8 linkers. The fusion complex was tested on loxF8R, loxF8R-5-zif268 (A), or loxF8R-5-zif268 (B) target sites. The test was performed at 200 ug/ml L-arabinose. Activity of the wild-type D7R is shown as a control. For (a), (b), (c) and (d) the upper band represents the unrecombined plasmid (line with two triangles), and the lower band represents the recombined plasmid (line with one triangle). M = Marker. The assays were performed three times (n = 3 biologically independent replicates).
