Evaluation of cationic dendrimer and lipid as transfection reagents of short RNAs for stem cell modification

摘要: Nowadays a large number of clinical trials suffer from lacking an efficient method for drug delivery into target cells with minimal side effects. Due to the great significance this issue in novel and effective therapies, more attempts are required order distinguish better conditions biomedical delivery. Since embryonic stem (ESCs) under scrutiny many new studies, development methods their genetical functional modifications is value. On other hand, application short nucleic acids therapeutic approaches increasing. In study efficiency small interfering RNA (siRNA) uptake two transfection reagents generation five polyamidoamine dendrimer (PAMAM G5) as cationic N-[1-(2,3-dioleoyloxy)]-N,N,N-trimethylammonium propane methyl-sulfate (DOTAP) lipid one commercially available reagent were evaluated mouse ESCs using flow cytometry. Prior cellular investigations; atomic force microscopy; gel electrophoresis; siRNA binding release assays; size zeta potential measurements utilized characterize physicochemical properties reagent-siRNA nano-complexes. The safety nano-complexes was subsequently assessed by MTT assay. Functional effects (complementary strand OCT4 transcript) mentioned analyzed quantitative real-time polymerase chain reaction (qPCR). Surprisingly DOTAP at higher molar ratios PAMAM lower could successfully knock down transcription relatively than commercial reagent. Our findings supported appropriate acid transfection. From point view, research helps allocation therapies.