Figure 6.

The sisiRNA design supports the silencing effects of chemically modified antisense strands. (A) Molecular structure of N2′-adamantylmethylcarbonyl 2′-amino-LNA-Thymine and N2′-pyren-1-ylmethyl 2′-amino-LNA-thymine. (B) Analyzing the effect of the sisiRNA design on the silencing efficiency of heavily modified antisense strands. The mean fluorescence of approximately 50 000 cells was measured by flow cytometry. The siRNA mismatch represents a siRNA that contains four mismatches to the EGFP target (Table 1). (C) The knockdown activity of the two siRNA duplex strands was assessed by measuring luciferase expression from reporter constructs containing either the target sequence for the sense or antisense strand of the EGFP specific duplex (light and dark gray bars, respectively). The reporter constructs are drawn above (not to scale). The experiment was performed in triplicate and for each reporter construct the firefly luciferase (Fluc)/renilla luciferase (Rluc) ratio was normalized to mismatch controls.