Affiliation:
1. School of Science RMIT University Melbourne Victoria 3000 Australia
2. Department of Chemical Engineering The University of Melbourne Parkville Victoria 3010 Australia
3. Dipartimento di Scienze e Tecnologie Chimiche Università degli Studi di Roma “Tor Vergata” Via della Ricerca Scientifica 1 Rome 00133 Italy
Abstract
AbstractNanoparticle‐mediated intracellular delivery of oligonucleotides is a complex phenomenon that depends on the architecture and the intracellular trafficking of the engineered nanoparticles. Unravelling the molecular arrangements of oligonucleotides within the nanoparticles as well as their intracellular behavior are essential for designing effective nucleic acid delivery systems. Herein, a simple and general strategy for probing the endosomal escape of nanoparticles carrying oligonucleotides in live cells is reported. A triplex‐forming oligonucleotide probe is designed to target the transcription factor, kappa‐light‐chain‐enhancer of activated B cells (NF‐κB), in the cytosol of cells and to transduce the binding into a fluorescent Förster resonance energy transfer (FRET) signal. The combined use of the triplex‐forming oligonucleotide probe and super‐resolution microscopy enables the elucidation of the morphology, intracellular localization, and endosomal escape of the oligonucleotide‐loaded nanoparticles on a molecular level and with nanoscale resolution. The co‐delivery of the FRET probe and mRNA in cells via lipid‐ and polymer‐ based nanoparticles allow simultaneous correlation of the endosomal escape properties of nanoparticles and gene expression efficiency.
Funder
Australian Research Council
H2020 Marie Skłodowska-Curie Actions
National Health and Medical Research Council
Subject
Electrochemistry,Condensed Matter Physics,Biomaterials,Electronic, Optical and Magnetic Materials