Self-assembling peptide–lipoplexes for substrate-mediated gene delivery

Abstract

The efficiency of biomaterial-based gene delivery is determined by the interaction of the material and the vector. For lipoplexes, surface immobilization has been used to transfect cells for applications such as cell arrays and model tissue formation through patterned transfection. Further increases in the delivery efficiency are limited by cellular internalization, which may be overcome by altering the material/vector interactions. In this report, we investigated the modification of the lipoplex physical properties through self-assembly with cationic peptides, and subsequently quantified cellular association, internalization and nuclear accumulation of DNA and transfection. Relative to lipid alone, peptide-lipoplexes enhanced transfection by up to 4.6-fold. The presence of the peptide in the lipoplex increased internalization efficiency by up to 4.5-fold, decreased the percentage of lysosomal DNA by 2.1-fold and increased the efficiency of nuclear accumulation by 3.0-fold. In addition, an analysis of internalization pathways for peptide-lipoplexes indicated a greater role of clathrin and caveolae-mediated endocytosis relative to macropinocytois, which was not observed for peptide-free lipoplexes. These results demonstrate peptide-induced enhancement of gene transfer by surface immobilization due to increased cellular internalization and nuclear accumulation, which has numerous applications ranging from cell-based assays to regenerative medicine.

Mesenchymal Stem Cells as a Novel Carrier for Targeted Delivery of Gene in Cancer Therapy Based on Nonviral Transfection

“…45 The transfection of the PEI 600 -Cyd/DNA complexes was examined in the presence of different endocytic inhibitors, including MβCD, to inhibit caveolae-mediated endocytosis, CPZ to inhibit clathrin-mediated endocytosis, and amiloride to inhibit macropinocytosisi. 46 The cell viability with PEI 600 -Cyd/DNA complexes and the inhibitors was found to be above 80%, indicating that these inhibitors are not toxic to MSCs ( Figure 2B). The cells treated with PEI 600 -Cyd/DNA complexes without adding any inhibitors were used as a control.…”

Studies of the physicochemical and structural properties of self-assembling cationic pyridine derivatives as gene delivery agents

“…Interest is growing towards the design of synthetic cationic lipid-like compounds as potential drug and gene delivery agents for transfer of genetic materials including high molecular weight DNA molecules into cells (Felgner et al, 1987) and for diagnostic applications (Marqués-Gallego and de Kroon, 2014). These lipid-like delivery systems are relatively non-toxic and non-immunogenic, moreover it is easy to use and produce them on a large scale (Al-Dosari and Gao, 2009;Guo and Huang, 2012;Lasic and Templeton, 1996;Margus et al, 2012;Rea et al, 2009).…”

SiRNA-loaded multi-shell nanoparticles incorporated into a multilayered film as a reservoir for gene silencing

“…Different systems were developed, e.g. polymers [8,9], liposomes [10,11], peptides [12,13], or inorganic nanoparticles like silica, magnetite, clay, gold, and calcium phosphate [14]. Such systems serve as a vector for active molecules which can pass through the cell membrane, escape from cytoplasmatic vesicles, and then reach their target in the cytoplasm.…”