High efficacy gold-KDEL peptide-siRNA nanoconstruct-mediated transfection in C2C12 myoblasts and myotubes

Abstract Gold nanoparticles (AuNP) were conjugated with cysteine terminated KDEL (Lys-Asp-Glu-Leu) peptide and siRNA directed against NADPH Oxidase 4 ( Nox4 ). Fluorescence microscopy analysis provided evidence of cytocellular retrograde transport pathways and sub-cellular colocalization of AuNP nan...

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Bibliographic Details
Published in:Nanomedicine 2014-02, Vol.10 (2), p.329-337
Main Authors: Acharya, Suresh, PhD, Hill, Rodney Allan, PhD
Format: Article
Language:English
Subjects:
Animals
Cell Line
Differentiation
Drug Delivery Systems
Endocytosis
Endoplasmic Reticulum - metabolism
Gene Silencing
Gold - chemistry
Gold nanoparticles
Golgi Apparatus - metabolism
Internal Medicine
KDEL
Metal Nanoparticles - chemistry
Mice
Muscle
Muscle Fibers, Skeletal - metabolism
Myoblasts - cytology
NADPH Oxidase 4
NADPH Oxidases - metabolism
Nanomedicine
Oligopeptides - chemistry
Peptide
Peptides - chemistry
Protein Sorting Signals
RNA, Small Interfering - chemistry
siRNA
Trafficking
Transfection
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Summary:Abstract Gold nanoparticles (AuNP) were conjugated with cysteine terminated KDEL (Lys-Asp-Glu-Leu) peptide and siRNA directed against NADPH Oxidase 4 ( Nox4 ). Fluorescence microscopy analysis provided evidence of cytocellular retrograde transport pathways and sub-cellular colocalization of AuNP nanoconstructs in both undifferentiated C2C12 myoblasts and differentiated C2C12 myotubes. The cellular trafficking of AuNP nanoconstructs in undifferentiated myoblasts suggests stable and efficient transfection of siRNA as demonstrated by colocalization of AuNP-delivered KDEL and siRNA. The cellular uptake of AuNP nanoconstructs was more efficient than Lipofectamine mediated transfection in differentiated myotubes ( P < 0.05) compared to undifferentiated myoblasts, suggesting that AuNP nanoconstructs provide an efficient platform for siRNA delivery to differentiated myotubes. The localization of these nanoconstructs in undifferentiated myoblasts suggests that most of the siRNA was localized in the endoplasmic reticulum (ER) with a minimal distribution in the Golgi bodies suggesting that the ER is a primary localization site for AuNP-KDEL mediated delivery of nanoconstructs. From the Clinical Editor This team of investigators demonstrate that a delivery system composed of gold nanoparticle and KDEL based siRNA is superior to lipofectamine in delivering siRNA in differentiated myoblasts, paving the way to gene silencing methods in these cells for future therapeutic exploitation.
ISSN:1549-9634
1549-9642
DOI:10.1016/j.nano.2013.07.015