Targeting reporter gene in integrated lentivirus for mutagenesis by the homing endonuclease Y2 I-AniI.

(a) Reporter (GFP) fluorescence and Y2 I-AniI expression (mCherry fluorescence) at the indicated days following transduction of the clonal reporter cell line T1H4S with lentiviral vector (moi of 2) expressing either the active enzyme Y2 I-AniI or the control inactive enzyme E148D I-AniI (as determined by mCherry expression), or control cells left untransduced (no enzyme, mCherry negative). This panel shows the data from one representative sample of a duplicate experiment. All cells were treated with 1 µM MG132 for 6 h prior to analysis. (b) Graphic representation of the data from the duplicate wells in the experiment depicted in Fig. 2a. Shown is the percent of the homing endonuclease-expressing cells (mCherry⁺ cell population) retaining reporter GFP fluorescence. The filled and open symbols correspond to the data from the duplicate wells. The percent of GFP⁺ cells was calculated as follows: {%GFP⁺ and mCherry⁺ cells x 100}/{Total % of mCherry⁺ cells}.