title: mPEG-PAMAM-G4 nucleic acid nanocomplexes: enhanced stability, RNase protection, and activity of splice switching oligomer and poly I:C RNA.
abstract: Dendrimer chemistries have virtually exploded in recent years with increasing interest in this class of polymers as gene delivery vehicles. An effective nucleic acid delivery vehicle must efficiently bind its cargo and form physically stable complexes. Most importantly, the nucleic acid must be protected in biological fluids and tissues, as RNA is extremely susceptible to nuclease degradation. Here, we characterized the association of nucleic acids with generation 4 PEGylated poly(amidoamine) dendrimer (mPEG-PAMAM-G4). We investigated the formation, size, and stability over time of the nanoplexes at various N/P ratios by gel shift and dynamic light scatter spectroscopy (DLS). Further characterization of the mPEG-PAMAM-G4/nucleic acid association was provided by atomic force microscopy (AFM) and by circular dichroism (CD). Importantly, mPEG-PAMAM-G4 complexation protected RNA from treatment with RNase A, degradation in serum, and various tissue homogenates. mPEG-PAMAM-G4 complexation also significantly enhanced the functional delivery of RNA in a novel engineered human melanoma cell line with splice-switching oligonucleotides (SSOs) targeting a recombinant luciferase transcript. mPEG-PAMAM-G4 triconjugates formed between gold nanoparticle (GNP) and particularly manganese oxide (MnO) nanorods, poly IC, an anticancer RNA, showed enhanced cancer-killing activity by an MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) cell viability assay.
authors.affiliation: Departments of ‡Biomedical Sciences, §Chemistry, and ∥Physics, Missouri State University , Springfield, Missouri 65897, United States.
authors.forename: Juan
authors.initials: J
authors.lastname: Reyes-Reveles
authors.forename: Reza
authors.initials: R
authors.lastname: Sedaghat-Herati
authors.forename: David R
authors.initials: DR
authors.lastname: Gilley
authors.forename: Ashley M
authors.initials: AM
authors.lastname: Schaeffer
authors.forename: Kartik C
authors.initials: KC
authors.lastname: Ghosh
authors.forename: Thomas D
authors.initials: TD
authors.lastname: Greene
authors.forename: Hannah E
authors.initials: HE
authors.lastname: Gann
authors.forename: Wesley A
authors.initials: WA
authors.lastname: Dowler
authors.forename: Stephen
authors.initials: S
authors.lastname: Kramer
authors.forename: John M
authors.initials: JM
authors.lastname: Dean
authors.forename: Robert K
authors.initials: RK
authors.lastname: Delong
mesh_terms: D017398:Alternative Splicing / Q000235:genetics*; D000818:Animals; D000970:Antineoplastic Agents / Q000378:metabolism; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D002942:Circular Dichroism; D050091:Dendrimers / Q000737:chemistry* / Q000494:pharmacology; D004337:Drug Carriers / Q000737:chemistry / Q000494:pharmacology; D005260:Female; D006801:Humans; D008545:Melanoma / Q000235:genetics / Q000473:pathology; D051379:Mice; D018625:Microscopy, Atomic Force; D015394:Molecular Structure; D049329:Nanostructures / Q000737:chemistry; D009757:Nylons / Q000737:chemistry* / Q000494:pharmacology; D009841:Oligonucleotides / Q000235:genetics* / Q000378:metabolism; D011070:Poly I-C / Q000235:genetics / Q000378:metabolism*; D011092:Polyethylene Glycols / Q000737:chemistry* / Q000494:pharmacology; D012313:RNA / Q000235:genetics / Q000378:metabolism; D020871:RNA Stability / Q000187:drug effects; D012259:Ribonuclease, Pancreatic / Q000378:metabolism*
chemical_list: D000970:Antineoplastic Agents; D050091:Dendrimers; D004337:Drug Carriers; D009757:Nylons; D009841:Oligonucleotides; C520550:PAMAM-G4; D011092:Polyethylene Glycols; D012313:RNA; D012259:Ribonuclease, Pancreatic; D011070:Poly I-C