1 research outputs found
mPEG-PAMAM-G4 Nucleic Acid Nanocomplexes: Enhanced Stability, RNase Protection, and Activity of Splice Switching Oligomer and Poly I:C RNA
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 polyÂ(amidoamine)
dendrimer (mPEG-PAMAM-G4). We investigated the formation, size, and
stability over time of the nanoplexes at various <i>N</i>/<i>P</i> 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