360. Magnetofection: A Versatile Approach for Messenger RNA Delivery

Abstract

Transcript therapy, using stabilized non-immunogenic messenger RNA (SNIM RNA), overcomes the potential risks of mutagenicity and immunogenicity, normally associated with DNA based gene delivery and recombinant protein therapy, respectively.Magnetofection, as a delivery system, has been shown to enhance DNA based viral and non-viral gene delivery. In this method, viral or non-viral gene carriers containing nucleic acids are associated with magnetic nanoparticles and a gradient magnetic field is applied to pull them on to the target cell surface.In this study, we established a magnetofection based protocol for enhanced mRNA delivery, using non-viral carriers. Firstly, we compared magnetofection, using different magnetic nanoparticles, to lipofection and polyfection, to find the best protocol for in vitro mRNA delivery. Then, the established protocol was generalized for co-transfection of up to three reporter SNIM RNAs (Luc, eGFP and RFP) in mono-culture of Primary Mouse Embryonic Fibroblasts (PMEF), or co-culture of PMEF and Porcine Fetal Fibroblast (PFF). In parallel, expression kinetics post magnetofection of d2eGFP SNIM RNA were determined.Magnetofection significantly improved transfection efficiency in mono culture as well as co-culture of applied cells. Using this protocol, co-transfection of different SNIM RNAs did not inhibit the transfection of every single SNIM RNA into PMEF or PFF cells. Moreover, rapid and higher peak of expression was observed with using magnetic nanoparticles.To conclude, magnetofection of mRNA presents itself as an efficient mRNA delivery system, thereby bringing transcript therapies a step closer to their clinical uses.