Working Title: ANTI-CANCER RNA NANOCONJUGATESInvestigators: Robert K. DeLong, Adam Wanekaya, and Kartik GhoshI. Abstract Nano-delivery of siRNA and RNA-based splice-site switching oligoribonucleotides(SSO) is an important new approach with chemotherapeutic potential against cancer. Here wewill focus on manufacturing RNA-bound nanoparticles (RNPs). For this purpose gold,manganese or zinc-based core nanoparticles (NPs) will be prepared and RNA attached viadelivery-enhancing materials (protamine or PAMAM dendrimer). In addition to a deliveryadvantage, this approach will greatly enhance RNA?s stability against chemical and enzymaticdegradation. Stabilization of the RNA by the RNPs will be confirmed by electrophoretic andliquid chromatography analysis after exposure to accelerated chemical or enzymaticdegradation assays. The RNPs will be further characterized by electron microscopy, UV,fluorescence and dynamic laser light scatter spectroscopy. Functional nuclear delivery of theRNPs will be evaluated in the Hela 705 splice-site switching assay by delivering a bound RNAoligonucleotide which overlaps the aberrant splice site, corrects splicing and switches-onLuciferase. Ultimately, as proof-of-principle to the important anti-cancer potential of thisapproach, biological activity of RNPs containing siRNA or SSO targeted against B-Raf?animportant model cancer target will be investigated.Keywords:SSO=splice-site switching oligonucleotide, RNP=RNA nanoparticle
This project focuses on nano-manufacturing nanoparticles of gold; manganese and zincwith RNA bound to them (RNPs). In preliminary experiments we have formed such RNPsattaching RNA via delivery enhancing materials (protamine or alternatively PAMAMdendrimer). Given these data and our previous experience with these nanomaterials incombination with DNA; here we will show the RNPs will provide the RNA resistance tochemical and enzymatic degradation and substantial delivery advantage. Finally as proof-of-principle for the chemotherapeutic potential of our approach here we will demonstrate thatRNPs can deliver siRNA or splice-site switching oligomers against B-Raf an important modelcancer target. Thus this proposal addresses a very important barrier for the success of RNA-derived chemotherapy; namely how to combine RNA with nanomaterials so as to retainstructure-function.
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