Figures & data
FIGURE 1 Locations of shRNAs targeting luciferase mRNA.
FIGURE 2 Lack of interferon induction by T7-transcribed shRNA. (a) The shRNA-luc constructs synthesized in these studies. We designed the pppGn (n = 0–3) associated with the 5′ end of the shRNA, which was transcribed by T7 RNA polymerase. We also designed the 5′-HOGn (n = 0–3) with the 5′ end of shRNA (removal of triphosphate by CIP). (b) The anti-luciferase activity of the pppGn (n = 0–3) associated with the 5′ end of shRNA-luc-3 in HeLaCD4+ cells. Firefly and Renilla luciferase activities were measured consecutively by using dual-luciferase assays (Promega) 48 h after transfection. (c) The residual amount of G associated with the 5′-end of the transcript is essential for the lack of interferon induction. HeLaCD4+ cells were transfected with 100 nM of pppGn (n = 0–3)-shRNAs or HOGn (n = 0–3)-shRNAs. The induced levels of IFN-β were determined by an ELISA.
