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Polymeric Nanocarriers for siRNA Delivery to Murine Macrophages
Author(s) -
Forbes Diane C.,
Peppas Nicholas A.
Publication year - 2014
Publication title -
macromolecular bioscience
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.924
H-Index - 105
eISSN - 1616-5195
pISSN - 1616-5187
DOI - 10.1002/mabi.201400027
Subject(s) - nanocarriers , internalization , chemistry , gene knockdown , nanoparticle , biophysics , polymer , flow cytometry , lipofectamine , förster resonance energy transfer , nanotechnology , fluorescence , materials science , cell , biochemistry , microbiology and biotechnology , apoptosis , organic chemistry , biology , recombinant dna , physics , quantum mechanics , gene , vector (molecular biology)
Abstract This work investigates the interactions of a polycationic nanocarrier with siRNA and with cells in order to better understand the capabilities and limitations of the carrier. The polycationic nanocarriers are cross‐linked copolymer nanoparticles synthesized in a single‐step reaction using ARGET ATRP (activators regenerated by electron transfer atom transfer radical polymerization). The polycationic nanocarriers efficiently bind siRNA for polymer/siRNA mass ratios less than 1. A method to prepare fluorescently labeled polycationic nanocarriers is presented. The fluorescently labeled polycationic nanocarriers are used to investigate cellular internalization with RAW264.7 murine macrophage cells. Flow cytometry demonstrates that the uptake increased with nanoparticle concentration and incubation time. Confocal microscopy confirmed internalization of fluorescently labeled nanoparticles. The investigation of siRNA‐induced knockdown demonstrates that higher concentrations of nanoparticles and siRNA are associated with increased knockdown. For the conditions tested in the knockdown experiments, the ARGET ATRP polycationic nanocarriers outperformed a commercially available Lipofectamine control.