Fig. 4: seRNA as a treatment approach against cancer and viral diseases.

seRNA_Δ3-5 (a) and seRNA (b) constructs with either eGFP or ca-Caspase3 were expressed in indicated cell types. Target-specific expression of eGFP (green) and cell viability was analyzed by light microscopy and flow cytometry, given in %. For (a) and (b) n = 3 or more. c Western blot analysis of HFF and U87 cells 24 h after transfection with seRNA-ca-Caspase3. Tubulin was used as an internal marker. n = 3. d Indicated glioblastoma cell lines were transfected with seRNA-ca-Caspase3 and tested for survival after 24 h. Toxicity is indicated in phase contrast. Numbers show relative killing efficiencies based on flow cytometry corrected by cell line-specific transfection efficiencies received with seRNA_Δ3-5−eGFP. n = 10 or more independent experiments. e seRNA-HBx expression plasmids were co-transfected with control and HBx expression plasmids. Maximal killing efficiencies were determined by using a constitutive ca-Caspase3 expression plasmid. 24 h after transfection, killing efficiencies were determined. n = 4 or more independent experiments. A student’s t test based significance level of 0.001 is displayed with three stars. Only positive relative killing efficiencies are illustrated in the figure. f Switch from plasmid-based expression to IVT seRNA. Indicated in vitro transcribed seRNAs were transfected into non-target (primary neurons) and U87 glioblastoma cells and analyzed for GFP expression and cell death. As a control, an IVT GFP-mRNA was used. Absolute GFP-transfection efficiencies and killing efficiencies are indicated based on flow cytometry analysis. n = 3 independent experiments. g Stably red nuclear labeled MCF-7 breast cancer cells and cytoplasmic GFP labeled human fibroblasts (HFF) without seRNA treatment and treatment for indicated times with seRNA-ca-Caspase3. Nuclear shapes are magnified for indicated areas. All scale bars = 200 µm except for (g) with 50 µm. n = 3.