Surface coating of siRNA–peptidomimetic nano-self-assemblies with anionic lipid bilayers: enhanced gene silencing and reduced adverse effects in vitro

Nanoscale ◽  
2015 ◽  
Vol 7 (46) ◽  
pp. 19687-19698 ◽  
Author(s):  
Xianghui Zeng ◽  
Anne Marit de Groot ◽  
Alice J. A. M. Sijts ◽  
Femke Broere ◽  
Erik Oude Blenke ◽  
...  
Keyword(s):  
Adverse Effects ◽  
Gene Silencing ◽  
Lipid Bilayers ◽  
Cellular Uptake ◽  
Surface Coating ◽  
Endosomal Escape ◽  
Anionic Lipid

Improved cellular uptake and endosomal escape of siRNA via surface coating of siRNA–peptidomimetic nanocomplexes with anionic lipid bilayers.

scholarly journals Design of New Polyaspartamide Copolymers for siRNA Delivery in Antiasthmatic Therapy

Pharmaceutics ◽  
2020 ◽  
Vol 12 (2) ◽  
pp. 89 ◽  
Author(s):  
Emanuela Fabiola Craparo ◽  
Salvatore Emanuele Drago ◽  
Nicolò Mauro ◽  
Gaetano Giammona ◽  
Gennara Cavallaro
Keyword(s):  
Epithelial Cells ◽  
Cellular Uptake ◽  
Intracellular Transport ◽  
Sirna Delivery ◽  
Weight Ratio ◽  
Endosomal Escape ◽  
Bronchial Epithelial ◽  
Vitro Experiment ◽  
In Vitro Experiment

Here, a novel protonable copolymer was realized for the production of polyplexes with a siRNA (inhibitor of STAT6 expression in asthma), with the aim of a pulmonary administration. The polycation was synthesized by derivatization of α,β-poly(N-2-hydroxyethyl)d,l-aspartamide (PHEA) with 1,2-Bis(3-aminopropylamino)ethane (bAPAE) in proper conditions to obtain a PHEA-g-bAPAE graft copolymer with a derivatization degree in amine (DDbAPAE%) equal to 35 mol%. The copolymer showed a proper buffering behavior, i.e., ranging between pH 5 and 7.4, to potentially give the endosomal escape of the obtained polycations. In effect, an in vitro experiment demonstrated the effect on biological membranes of the copolymer on bronchial epithelial cells (16-HBE) strongly dependent on the pH of the medium, i.e., higher at pH 5. bAPAE-based copolymers were further obtained with an increasing pegylation degree, i.e., equal to 1.9, 2.7, and 4.4 mol%, respectively. All the obtained copolymers were able to complex siRNA at a N/P ratio that decreases as the pegylation degree increases. At the same time, the tendency of polyplexes to aggregate and the capability to interact with mucin also decreases as the pegylation in the copolymer increases. Gene silencing experiments on 16-HBE showed that these copolymers have a significant role in improving the intracellular transport of naked siRNA, where the presence of PEG does not seem to hinder the cellular uptake of polyplexes. The latter obtained at polymer/siRNA weight ratio (R) equal to 10 with PHEA-g-PEG(C)-g-bAPAE also seems to be not susceptible to the presence of mucin, avoiding the polyanionic exchange of complexed siRNA, thus showing adequate behavior to be used as an effective vector for siRNA.

scholarly journals Novel fusion peptide‐mediated siRNA delivery using self‐assembled nanocomplex

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Yeong Chae Ryu ◽  
Kyung Ah Kim ◽  
Byoung Choul Kim ◽  
Hui-Min David Wang ◽  
Byeong Hee Hwang
Keyword(s):  
Gene Silencing ◽  
Cellular Uptake ◽  
Viral Infections ◽  
Immune Cell ◽  
Sirna Delivery ◽  
Cellular Membrane ◽  
Fusion Peptides ◽  
Self Assembled

Abstract Background Gene silencing using siRNA can be a new potent strategy to treat many incurable diseases at the genetic level, including cancer and viral infections. Treatments using siRNA essentially requires an efficient and safe method of delivering siRNA into cells while maintaining its stability. Thus, we designed novel synergistic fusion peptides, i.e., SPACE and oligoarginine. Results Among the novel fusion peptides and siRNAs, nanocomplexes have enhanced cellular uptake and gene silencing effect in vitro and improved retention and gene silencing effects of siRNAs in vivo. Oligoarginine could attract siRNAs electrostatically to form stable and self-assembled nanocomplexes, and the SPACE peptide could interact with the cellular membrane via hydrogen bonding. Therefore, nanocomplexes using fusion peptides showed improved and evident cellular uptake and gene silencing of glyceraldehyde 3-phosphate dehydrogenase (GAPDH) via the lipid raft-mediated endocytosis pathway, especially to the HDFn cells of the skin, and all of the fusion peptides were biocompatible. Also, intratumorally injected nanocomplexes had increased retention time of siRNAs at the site of the tumor. Finally, nanocomplexes demonstrated significant in vivo gene silencing effect without overt tissue damage and immune cell infiltration. Conclusions The new nanocomplex strategy could become a safe and efficient platform for the delivery of siRNAs into cells and tissues to treat various target diseases through gene silencing.

A pH-sensitive fusogenic peptide facilitates endosomal escape and greatly enhances the gene silencing of siRNA-containing nanoparticles in vitro and in vivo

2009 ◽  
Vol 139 (2) ◽  
pp. 127-132 ◽  
Author(s):  
Hiroto Hatakeyama ◽  
Erika Ito ◽  
Hidetaka Akita ◽  
Motoi Oishi ◽  
Yukio Nagasaki ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Ph Sensitive ◽  
Endosomal Escape

scholarly journals Cationic liposome–nucleic acid nanoparticle assemblies with applications in gene delivery and gene silencing

2016 ◽  
Vol 374 (2072) ◽  
pp. 20150129 ◽  
Author(s):  
Ramsey N. Majzoub ◽  
Kai K. Ewert ◽  
Cyrus R. Safinya
Keyword(s):  
Nucleic Acid ◽  
Gene Delivery ◽  
Gene Silencing ◽  
Liquid Crystalline ◽  
Cationic Liposome ◽  
Endosomal Escape ◽  
Cubic Phase ◽  
Nucleic Acid Therapeutics

Cationic liposomes (CLs) are synthetic carriers of nucleic acids in gene delivery and gene silencing therapeutics. The introduction will describe the structures of distinct liquid crystalline phases of CL–nucleic acid complexes, which were revealed in earlier synchrotron small-angle X-ray scattering experiments. When mixed with plasmid DNA, CLs containing lipids with distinct shapes spontaneously undergo topological transitions into self-assembled lamellar, inverse hexagonal, and hexagonal CL–DNA phases. CLs containing cubic phase lipids are observed to readily mix with short interfering RNA (siRNA) molecules creating double gyroid CL–siRNA phases for gene silencing. Custom synthesis of multivalent lipids and a range of novel polyethylene glycol (PEG)-lipids with attached targeting ligands and hydrolysable moieties have led to functionalized equilibrium nanoparticles (NPs) optimized for cell targeting, uptake or endosomal escape. Very recent experiments are described with surface-functionalized PEGylated CL–DNA NPs, including fluorescence microscopy colocalization with members of the Rab family of GTPases, which directly reveal interactions with cell membranes and NP pathways. In vitro optimization of CL–DNA and CL–siRNA NPs with relevant primary cancer cells is expected to impact nucleic acid therapeutics in vivo . This article is part of the themed issue ‘Soft interfacial materials: from fundamentals to formulation’.

scholarly journals Novel Fusion Peptide-Mediated siRNA Delivery Using Self-Assembled Nanocomplex

2020 ◽  
Author(s):  
Yeong Chae Ryu ◽  
Kyungah Kim ◽  
Byoung Choul Kim ◽  
Hui-Min David Wang ◽  
Byeong Hee Hwang
Keyword(s):  
Gene Therapy ◽  
Gene Silencing ◽  
Cellular Uptake ◽  
Viral Infections ◽  
Sirna Delivery ◽  
Cellular Membrane ◽  
Fusion Peptides ◽  
Self Assembled

Abstract Background: Gene therapy using siRNA can be a new potent strategy to treat many incurable diseases at the genetic level, including cancer and viral infections. Treatments using siRNA essentially requires an efficient and safe method of delivering siRNA into cells while maintaining its stability. Thus, we designed novel synergistic fusion peptides, i.e., SPACE and oligoarginine.Results: Among the novel fusion peptides and siRNAs, nanocomplexes have outstanding cellular uptake and gene silencing effect in vitro and high stability and retention effect of siRNAs in vivo. Oligo arginine could attract siRNAs electrostatically to form stable and self-assembled nanocomplexes, and the SPACE peptide could interact with the cellular membrane via hydrogen bonding. Therefore, nanocomplexes using fusion peptides showed improved and evident cellular uptake and gene silencing of Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) via the lipid raft-mediated endocytosis pathway, especially to the HDFn cells of the skin, and all of the fusion peptides were biocompatible. Also, intratumorally injected nanocomplexes had increased stability and retention of siRNAs at the site of the tumor. Conclusions: The new nanocomplex strategy could become a safe and efficient platform for the delivery of siRNAs into cells and tissues to treat various target diseases through gene therapy.

Surface modified PMMA nanoparticles with tunable drug release and cellular uptake

RSC Advances ◽  
2014 ◽  
Vol 4 (84) ◽  
pp. 44472-44479 ◽  
Author(s):  
Ridhima Juneja ◽  
Indrajit Roy
Keyword(s):  
Drug Release ◽  
Cellular Uptake ◽  
Surface Coating ◽  
Uptake Efficiency ◽  
Surface Modified ◽  
Cellular Uptake Efficiency

Differentialin vitrocellular uptake efficiency of fluorophore-loaded PMMA nanoparticles, with (a) different size, and (b) different surface coating.

scholarly journals Novel fusion peptide-mediated siRNA delivery using self-assembled nanocomplex

2021 ◽  
Author(s):  
Yeong Chae Ryu ◽  
Kyungah Kim ◽  
Byoung Choul Kim ◽  
Hui-Min David Wang ◽  
BYEONG HEE HWANG
Keyword(s):  
Gene Silencing ◽  
Cellular Uptake ◽  
Viral Infections ◽  
Sirna Delivery ◽  
Cellular Membrane ◽  
Fusion Peptides ◽  
Genetic Level ◽  
Self Assembled

Abstract Background: Gene silencing using siRNA can be a new potent strategy to treat many incurable diseases at the genetic level, including cancer and viral infections. Treatments using siRNA essentially requires an efficient and safe method of delivering siRNA into cells while maintaining its stability. Thus, we designed novel synergistic fusion peptides, i.e., SPACE and oligoarginine.Results: Among the novel fusion peptides and siRNAs, nanocomplexes have enhanced cellular uptake and gene silencing effect in vitro and improved retention and gene silencing effects of siRNAs in vivo. Oligoarginine could attract siRNAs electrostatically to form stable and self-assembled nanocomplexes, and the SPACE peptide could interact with the cellular membrane via hydrogen bonding. Therefore, nanocomplexes using fusion peptides showed improved and evident cellular uptake and gene silencing of Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) via the lipid raft-mediated endocytosis pathway, especially to the HDFn cells of the skin, and all of the fusion peptides were biocompatible. Also, intratumorally injected nanocomplexes had increased retention time of siRNAs at the site of the tumor. Finally, nanocomplexes demonstrated significant in vivo gene silencing effect without immunogenicity.Conclusions: The new nanocomplex strategy could become a safe and efficient platform for the delivery of siRNAs into cells and tissues to treat various target diseases through gene silencing.

Using FRET to Measure the Time it Takes for a Cell to Destroy a Virus

2019 ◽  
Author(s):  
Candace E. Benjamin ◽  
Zhuo Chen ◽  
Olivia Brohlin ◽  
Hamilton Lee ◽  
Stefanie Boyd ◽  
...  
Keyword(s):  
Cellular Uptake ◽  
Rhodamine B ◽  
Virus Like Particle ◽  
A Cell ◽  
Viral Nanoparticles ◽  
The Stability ◽  
Open Question ◽  
Viral Surface ◽  
Fret Pair

<div><div><div><p>The emergence of viral nanotechnology over the preceding two decades has created a number of intellectually captivating possible translational applications; however, the in vitro fate of the viral nanoparticles in cells remains an open question. Herein, we investigate the stability and lifetime of virus-like particle (VLP) Qβ - a representative and popular VLP for several applications - following cellular uptake. By exploiting the available functional handles on the viral surface, we have orthogonally installed the known FRET pair, FITC and Rhodamine B, to gain insight of the particle’s behavior in vitro. Based on these data, we believe VLPs undergo aggregation in addition to the anticipated proteolysis within a few hours of cellular uptake.</p></div></div></div>

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