Different dsRNA expression strategies have different RNAi potential.

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Different dsRNA expression strategies have different RNAi potential. Different dsRNA expression strategies have different RNAi potential. (A) Schematic depiction of relevant expressed portions of plasmids used for dsRNA expression and types of dsRNAs produced. (B) Impact of Mos dsRNA on RL-Mos in normal and Pkr−/− 3T3 cells. 3T3 cells were transfected with FL, RL-Mos reporters, and a Mos dsRNA-expressing plasmid or its Lin28 counterpart. Relative Renilla luciferase activity (RL-Mos/FL) in Mos dsRNA-expressing cells is shown relative to that in Lin28 dsRNA-expressing cells for each dsRNA type and genotype. The experiment was performed twice in triplicates. Error bars = SD. (C) Impact of Lin28 dsRNA on RL-Lin28 in normal and PKR−/− 3T3 cells. 3T3 cells were transfected with FL, RL-Lin28, and Lin28 dsRNA-expressing plasmid or its Mos counterpart. Relative Renilla luciferase activity (RL-Lin28/FL) in Lin28 dsRNA-expressing cells is shown relative to that in Mos dsRNA-expressing cells for each dsRNA type and genotype. The experiment was performed twice in triplicates. Error bars = SD. Note that loss of Pkr has a small, mostly statistically insignificant, positive effect on RNAi in some cases, pol III–driven Lin28 hairpin is inducing strong RNAi effect regardless of the presence or absence of PKR. Tomas Demeter et al. LSA 2019;2:e201800289 © 2019 Demeter et al.