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

By Martine Aubert (356243), Byoung Y. Ryu (356244), Lindsey Banks (356245), David J. Rawlings (356246), Andrew M. Scharenberg (75184) and Keith R. Jerome (356247)

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<sup>+</sup> cell population) retaining reporter GFP fluorescence. The filled and open symbols correspond to the data from the duplicate wells. The percent of GFP<sup>+</sup> cells was calculated as follows: {%GFP<sup>+</sup> and mCherry<sup>+</sup> cells x 100}/{Total % of mCherry<sup>+</sup> cells}.</p

Topics: Genetics, Biotechnology, Infectious Diseases, Virology, lentivirus, mutagenesis, homing, endonuclease, y2
Year: 2013
DOI identifier: 10.1371/journal.pone.0016825.g002
OAI identifier: oai:figshare.com:article/469712
Provided by: FigShare
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