Hydrolytic charge‐reversal of PEG ylated polyplexes enhances intracellular un‐packaging and activity of si RNA

Autor: Corban Swain, Kameron V. Kilchrist, Craig L. Duvall, Thomas A. Werfel, Brian C. Evans, Christopher E. Nelson, Martina Miteva
Rok vydání: 2016
Předmět:
inorganic chemicals
Materials science
Cell Survival
Carboxylic Acids
Biomedical Engineering
02 engineering and technology
010402 general chemistry
Methacrylate
Hemolysis
01 natural sciences
Article
Polyethylene Glycols
Biomaterials
chemistry.chemical_compound
Cell Line
Tumor
mental disorders
PEG ratio
Fluorescence Resonance Energy Transfer
Humans
Gene silencing
Amines
RNA
Small Interfering
Cytotoxicity
health care economics and organizations
Microscopy
Confocal
Hydrolysis
technology
industry
and agriculture
Metals and Alloys
Cationic polymerization
Raft
respiratory system
021001 nanoscience & nanotechnology
Nanostructures
0104 chemical sciences
chemistry
Ceramics and Composites
Biophysics
Methacrylates
RNA Interference
0210 nano-technology
Ethylene glycol
Intracellular
Biomedical engineering
Zdroj: Journal of Biomedical Materials Research Part A. 104:917-927
ISSN: 1552-4965
1549-3296
DOI: 10.1002/jbm.a.35629
Popis: Hydrolytically degrading nano-polyplexes (HDG-NPs) that reverse charge through conversion of tertiary amines to carboxylic acids were investigated to improve intracellular un-packaging of siRNA and target gene silencing compared to a non-degradable analog (non-HDG-NPs). Both NP types comprised reversible addition-fragmentation chain-transfer (RAFT) synthesized diblock copolymers of a poly(ethylene glycol) (PEG) corona-forming block and a cationic block for nucleic acid packaging that incorporated butyl methacrylate (BMA) and either dimethylaminoethyl methacrylate (DMAEMA, non-HDG-NPs) or dimethylaminoethyl acrylate (DMAEA, HDG-NPs). HDG-NPs decreased significantly in size and released significantly more siRNA (∼40%) than non-HDG-NPs after 24 h in aqueous solution. While both HDG-NPs and non-HDG-NPs had comparable uptake and cytotoxicity up to 150 nM siRNA doses, HDG-NPs achieved significantly higher target gene silencing of the model gene luciferase in vitro. High resolution FRET confocal microscopy was used to monitor the intracellular un-packaging of siRNA. Non-HDG-NPs had significantly higher FRET efficiency than HDG-NPs, indicating that siRNA delivered from HDG-NPs was more fully un-packaged and therefore had improved intracellular bioavailability.
Databáze: OpenAIRE
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