Targeting reporter gene in integrated lentivirus for mutagenesis by the homing endonuclease Y2 I-<i>Ani</i>I.

Abstract

<p>(<b>a</b>) Reporter (GFP) fluorescence and Y2 I-<i>Ani</i>I 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-<i>Ani</i>I or the control inactive enzyme E148D I-<i>Ani</i>I (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>b</b>) Graphic representation of the data from the duplicate wells in the experiment depicted in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0016825#pone-0016825-g002" target="_blank">Fig. 2a</a>. 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}.</p