Abstract
Messenger RNA (mRNA) is not an attractive candidate for gene therapy due to its instability and has therefore received little attention. Recent studies show the advantage of mRNA over DNA, especially in cancer immunotherapy and vaccine development. This study aimed to formulate folic-acid-(FA)-modified, poly-amidoamine-generation-5 (PAMAM G5D)-grafted gold nanoparticles (AuNPs) and to evaluate their cytotoxicity and transgene expression using the luciferase reporter gene (FLuc-mRNA) in vitro. Nanocomplexes were spherical and of favorable size. Nanocomplexes at optimum nanoparticle:mRNA (w/w) binding ratios showed good protection of the bound mRNA against nucleases and were well tolerated in all cell lines. Transgene expression was significantly (p < 0.0001) higher with FA-targeted, dendrimer-grafted AuNPs (Au:G5D:FA) in FA receptors overexpressing MCF-7 and KB cells compared to the G5D and G5D:FA NPs, decreasing significantly (p < 0.01) in the presence of excess competing FA ligand, which confirmed nanocomplex uptake via receptor mediation. Overall, transgene expression of the Au:G5D and Au:G5D:FA nanocomplexes exceeded that of G5D and G5D:FA nanocomplexes, indicating the pivotal role played by the inclusion of the AuNP delivery system. The favorable properties imparted by the AuNPs potentiated an increased level of luciferase gene expression.
Highlights
Over the years, non-viral gene delivery modalities based on plasmid DNAwere extensively evaluated in vitro as potential treatments of inherited diseases [1]
Were extensively evaluated in vitro as potential treatments of inherited diseases [1]. Their failure to demonstrate potency at a clinical level due to their inability to bypass hurdles posed by the nuclear membrane of non-dividing cells and immunogenic responses of cytosine-phosphate-guanine (CpG) motifs contained by unmethylated DNA has aroused interest in using Messenger RNA (mRNA) instead of plasmid DNA (pDNA) [2,3]
The covalent attachment of the Folic Acid (FA) onto the surface of NPs is known by its absorption maxima at 280 nm with a saddle point at 360 nm [34,35] (Figure 1A), corresponding to the absorption peak of Au:G5D:FA observed at 287 nm
Summary
Were extensively evaluated in vitro as potential treatments of inherited diseases [1] Their failure to demonstrate potency at a clinical level due to their inability to bypass hurdles posed by the nuclear membrane of non-dividing cells and immunogenic responses of cytosine-phosphate-guanine (CpG) motifs contained by unmethylated DNA has aroused interest in using mRNA instead of pDNA [2,3]. The recent interest in mRNA-based systems is due to the pharmaceutical safety advantages demonstrated over their pDNA-based counterparts These include, first, the ease of mRNA to be formulated into an efficient therapeutic agent since it does not require the incorporation of promoters and terminators such as pDNA. MRNA can transfect non-dividing cells, and its inability
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
