Fig. 4.
In vitro translations with luciferase reporter assay to assess P-PMO 5UP1's effect on variable target sequences. (A) DNA versions of the complementary target sequence of 5UP1 and five variants of the target sequence with one to four mismatches (MM) each were cloned upstream of luciferase in reporter plasmids. RNA was in vitro transcribed and in vitro translation reactions were conducted to measure luciferase translation by the six RNAs in the presence of various concentrations of P-PMO 5UP1. EC50s are shown and represent the effective concentration of a compound resulting in a 50% reduction in luciferase signal compared with mock treatment controls. (B) Relative percentages of inhibition were calculated in comparison with mock treatment controls. Note the decreasing level of inhibition of luciferase production caused by P-PMO 5UP1 against RNAs with which it has an increasing number of mispairs.