Female C57/BL6 mice were injected with LNPs containing siRNA or dsiRNA. Mice were dosed with 0.6, 0.1, or 0.02 mg/kg for FVII-targeting compounds and controls. The dose of dsiRNA was adjusted such that the amount of 21-mer produced by Dicer cleavage in vivo (assuming 100% accurate and complete cleavage) was equal to the amount of canonical siRNA administered. For PTEN-targeting compounds and controls, mice received 0.4, 0.1, or 0.02 mg/kg with dosing adjusted for dsiRNA as described. Blood (FVII) or liver (PTEN) was collected 2 d after injection. Factor VII protein was quantified by chromogenic assay, while PTEN mRNA was quantified by branched DNA. Data were normalized to Luciferase-targeted siRNA control. (A,B) ED50 values were determined by nonlinear regression of Day 2 activity data. Log-transformed ED50 data were compared by two-way ANOVA (structure × dose). (*) p ≤ 0.05. (C–F) Dose-response data obtained 2 d after injection. Canonical leads with dsiRNA matches (C,E), as well as dsiRNA leads with canonical siRNA matches (D,F), are shown. Modifications are (UU) unmodified/unmodified, (HL) heavy/light, (HA) heavy/alternating, and (UA) unmodified/alternating.
