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. 2015 Apr 14;23(6):1055–1065. doi: 10.1038/mt.2015.41

Figure 1.

Figure 1

ZFN targeting of intronic GAA repeats in the FXN gene. (a) Diagram of the FXN gene ZFN editing strategy. Exact location of the UP- and DN-ZFNs is shown relative to the proximal and distal end of the GAA repeat region. Approximate locations of the PCR primers (UP-F/R and DN-F/R) used to amplify ZFN targeted region are indicated. Exact locations and sequences of the primers and ZFNs are shown in Supplementary Figure S1. (b) Simultaneous cleavage by UP- and DN-ZFNs. K562 cells were cotransfected with UP/DN-ZFN mRNAs. In K562 cells harboring the short GAA tract, editing of intron 1 of the FXN gene leads to the release of ~1.2 kbp fragment. Deletion can be detected by PCR using UP-F and DN-R primers. Amplification of the non-GAA edited allele results in a 1.6-kbp fragment (lane C) while amplification of ZFN-edited DNA results in a 0.4-kbp fragment (lane UP-ZFN, DN-ZFN). L—Hyperladder I (BioLine). (c) Schematic illustrating ZFN-mediated editing of the FXN locus.