Effect of distance of the intronic regulatory region to the natural 5’ splice site.

<p>The upper panel shows the location and sequence of the spacer introduced in the pcDNA3.1 wild type minigene between the natural and the cryptic splice sites (GT underlined). The gel shows the results after transfection of the wild-type (WT) minigene and the constructs with 1, 3 or 6 spacers in Hep3B cells. On the right of the gel is the schematic drawing showing the identity of the bands verified by sequence analysis, that showed usage of both the natural and the cryptic splice site (at +24 with 1 spacer, at +36 with 3 spacers and at +54 with 6 spacers). The estimated percentage of exon inclusion is shown below each lane.