Figure 1.

In vitro-transcribed RNA is immunogenic and contains dsRNA contaminants. (A) 200 ng of in vitro transcripts encoding mEPO and containing the indicated modified nucleosides were blotted and analyzed with K1 and J2 dsRNA-specific mAbs. The dsRNA positive control contained a 328 bp long dsRNA (25 ng). (B) DCs were treated with Lipofectin-complexed Renilla luciferase (T7TSRenA₃₀), firefly and Metridia luciferases (T7TSLucA₃₀, T7TSMetlucA₃₀), and mEPO (TEVmEPOA₅₁) mRNAs. TNF-α levels were measured in the supernatants at 24 h. (C) DCs were treated with TransIT-complexed in vitro transcripts encoding Renilla and firefly luciferases (T7TSRenA₃₀, T7TSLucA₃₀), eGFP (TEVeGFPA₅₁) and mEPO (TEVmEPOA₅₁). IFN-α levels were measured in the supernatants at 24 h. Error bars are standard error of the mean. Data shown is from one experiment that is representative of 3–6.