Supplementary Figure 1: Validation of the length for target sequence.

(a and b) To validate the binding affinity of TALEs, we designed 4 different TALEs against 13, 15, 18, and 20 nt of MajSat sequence and fused them with the VP64 activation domain (B13, B15, B18, and B20, respectively). The firefly luciferase reporter plasmid contains tandem repeats of the binding sequence upstream of a minimal CMV promoter. (b) The indicated TALE-VP64 fusions and the reporter plasmid were transfected to 293T cells. Relative luciferase activity normalized to Renilla luciferase activity is shown. NC indicates mock transfection control. The mean ± s.d. of three independent biological replicates is shown. We found progressive increase of luciferase activity with the length of the target sequence, in agreement with a previous report ⁸. (c) The indicated TALE-mClover constructs were transfected in mouse ES cells. Confocal images of TALE-mClover and DAPI are shown. line-profiles (RGB profile) of fluorescence intensities for DAPI (black) and mClover (green) from the A-B line drawn in images are shown in right. Note that B13 displays slightly higher background signal in nucleoplasm contrasting to other TALEs. Scale bar, 2 μm.