A self-assembling amphiphilic peptide nanoparticle for the efficient entrapment of DNA cargoes up to 100 nucleotides in length.

Abstract - To overcome the low efficiency and cytotoxicity associated with most non-viral DNA delivery systems we developed a purely peptidic self-assembling system that is able to entrap single- and double-stranded DNA of up to 100 nucleotides in length. (HR)3gT peptide design consists of a hydrophilic domain prone to undergo electrostatic interactions with DNA cargo, and a hydrophobic domain at a ratio that promotes the self-assembly into multi-compartment micellar nanoparticles (MCM-NPs). #Self-assembled (HR)3gT MCM-NPs range between 100 to 180 nm which is conducive to a rapid and efficient uptake by cells. (HR)3gT MCM-NPs had no adverse effects on HeLa cell viability. In addition, they exhibit long-term structural stability at 4 °C but at 37 °C, the multi-micellar organization disassembles overtime which demonstrates their thermo-responsiveness. The comparison of (HR)3gT to a shorter, less charged H3gT peptide indicates that the additional arginine residues result in the incorporation of longer DNA segments, an improved DNA entrapment efficiency and an increase cellular uptake. Our unique non-viral system for DNA delivery sets the stage for developing amphiphilic peptide nanoparticles as candidates for future systemic gene delivery.

1 Department of Chemistry, University of Basel, Mattenstrasse 24a, 4058 Basel, Switzerland. cornelia.palivan@unibas.ch and Department of Biosystems Science and Engineering, ETH Zurich, Mattenstrasse 26, 4058 Basel, Switzerland. kobi.benenson@bsse.ethz.ch.
2 Department of Chemistry, University of Basel, Mattenstrasse 24a, 4058 Basel, Switzerland. cornelia.palivan@unibas.ch.
3 Department of Biosystems Science and Engineering, ETH Zurich, Mattenstrasse 26, 4058 Basel, Switzerland. kobi.benenson@bsse.ethz.ch.

More:
Solvents and reagents
Peptide synthesis and purification
Self-assembly of peptide MCM-NPs
Characterization of DNA-free and DNA-loaded peptide MCM-NPs
Thermo-responsiveness of DNA-free and DNA-loaded peptide MCM-NPs
Characterization of DNA-free and DNA-loaded peptide MCM-NPs
https://pubs.rsc.org/en/content/articlelanding/202...