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. 2017 Mar 8;8(24):38239–38250. doi: 10.18632/oncotarget.16013

Figure 1. Design and optimisation of vectors.

Figure 1

A. Schematic representation of vector design showing the AluYa8 element flanked with the HSV-TK gene (grey boxes) followed by a pA2 cleavage site and GFP (green box). Both HSV-TK and GFP are on the negative strand with the CMV promoter (yellow box). The HSV-TK coding sequence is interrupted by the presence of an autocatalytic Tetrahymena (Tet) intron (light blue box), which can be spliced out of the transposition RNA intermediate (middle). The predicted structure of the resulting de novo Alu insertion following splicing and reverse transcription is shown (bottom). B. Table summarizing the differences between the constructs. hGHpa is human growth hormone poly A. C. Histograms showing the percentage of DAPI stained cells with GFP signal for each construct. 5×104 HEK293T/17 cells were grown in a 12-well plate and transfected with 1μg DNA using FuGENE® (Promega) (DNA/FuGENE ratio 1/6). GFP signal was measured at 24, 48 and 72 hours post transfection using CellInsight, (Cellomics, Thermofisher) and the data were analysed using HCS Studio.