A versatile reducible polycation-based system for efficient delivery of a broad range of nucleic acids

Abstract

Synthetic vectors based on reducible polycations consisting of histidine and polylysine residues (HIS RPCs) were evaluated for their ability to deliver nucleic acids. Initial experiments showed that RPC-based vectors with at least 70% histidine content mediated efficient levels of gene transfer without requirement for the endosomolytic agent chloroquine. Significant gene transfer was observed in a range of cell types achieving up to a 5-fold increase in the percentage of transfected cells compared to 25 kDa PEI, a gold standard synthetic vector. In contrast to 25 kDa PEI, HIS RPCs also mediated efficient transfer of other nucleic acids, including mRNA encoding green fluorescent protein in PC-3 cells and siRNA directed against the neurotrophin receptor p75NTR in post-mitotic cultures of rat dorsal root ganglion cell neurons. Experiments to elevate intracellular glutathione and linear profiling of cell images captured by multiphoton fluorescent microscopy highlighted that parameters such as the molecular weight and rate of cleavage of HIS RPCs were important factors in determining transfection activity. Altogether, these results demonstrate that HIS RPCs represent a novel and versatile type of vector that can be used for efficient cytoplasmic delivery of a broad range of nucleic acids. This should enable different or a combination of therapeutic strategies to be evaluated using a single type of polycation-based vector.

Receptor-mediated gene delivery using polyethylenimine (PEI) coupled with polypeptides targeting FGF receptors on cells surface

“…Since Boussif et al(1995) hope for non-viral vector in gene delivery, PEI has become the golden criteria in the field of non-viral gene transfer vectors (Read et al, 2005b). To make PEI more suitable for gene therapy, modifying PEI with ligands is one of the useful strategies, targeting specially to the receptors on cell membrane to improve the targeting ability of PEI.…”

Bioreducible Cationic Polymers for Gene Transfection

“…Also, different dithiol-bearing groups, e.g. nuclear localization sequences comprising two cysteine residues, can be incorporated in the oxidation reaction, giving rise to disulfide-containing copolymers with multiple functionalities (Read et al, 2005).…”

Reducible poly(amido ethylenediamine) for hypoxia-inducible VEGF delivery

“…D,L -Buthionine sulfoxamine (BSO), a known GSH inhibitor, was used as previously described by Read et al [22]. H9C2 cells were plated 48 hrs prior to BSO treatment at 1.0 × 10 5 cells/ well in 6-well plates.…”

“…Seymour and coworkers showed that the intracellular levels of glutathione (GSH) affected nucleic acid delivery of their reducible polycations [22]. Previous work in mouse fibroblast cells also showed that upon internalization of SS-PAEI complexed with fluorescent labeled pDNA, complexes showed a wider dispersion of pDNA as compared to a non-reducible bPEI control.…”