engleski

RNA interference in drug development: nanotechnology and cell viability

RNA interference (RNAi) is an endogenously present regulatory mechanism in most eukaryote cells. It involves the destruction of messenger RNA (mRNA) upon interaction with homologous double stranded RNA (dsRNA), which is also known as small interfering RNA (siRNA) and can be exogenously introduced into the cells. This mechanism has shown great potential for drug development. Different methods for delivering siRNA into cells have been developed. Electroporation enables the delivery of naked, negatively charged, siRNA into the cell. However, during electroporation, cells are exposed to a high-voltage pulse, which can cause enormous reduction in cell viability. As opposed to electroporation, the complexing of siRNA in NP with net positive surface charge ought to enable more efficient delivery of siRNA into the cell with less extent of cytotoxicity. In our study, aqueous solutions of chitosan (in the form of glutamate salt (<200kDa)) pentasodium tripolyphosphate and siRNA were used for NP preparation. RPMI medium was used for Jurkat cell line growth. Hank's buffered salt solution (HBSS) of various pH was used for 1h or 2h incubation of NP with cells. The Eppendorf Hypoosmolar electroporation buffer was used for electroporation. Mean particle size of prepared siRNA-NPs was in the nano-range (~300-400nm). Surface charge of siRNA-NP was positive when NPs were incubated in HBSS with pH<6, 5. We examined the citotoxicity of prepared siRNA-NPs using MTT and lactate dehydrogenase (LDH) assays and found that cytotoxicity had different yield for cells incubated in the HBSS buffer of various pH.

RNA interference; siRNA; nanoparticles; chitosan

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