Improved transfection efficiency of an aliphatic lipid substituted 2 kDa polyethylenimine is attributed to enhanced nuclear association and uptake in rat bone marrow stromal cell

Hsu, Charlie Yu Ming; Hendzel, Michael J.; Uludağ, Hasan

doi:10.1002/jgm.1526

Cited by 38 publications

(19 citation statements)

References 36 publications

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“…The presence of 10% serum, which was avoided by most studies, may explain the greater size of polyplexes observed in this work. In one study, where the size measurements of PEI polyplexes were reported in 1% serum, the observed sizes (210-663 nm) were in line with the current study at similar DNA and polymer concentrations [32].…”

Section: Sizes Of Pei-peg Polyplexessupporting

confidence: 90%

“…The sources of all plasmids and tissue culture reagents were described previously [32]. SYBR Green I was purchased from Invitrogen (Carlsbad, CA).…”

Section: Methodsmentioning

confidence: 99%

“…After 5 h, the medium was replaced, and cells were processed for flow cytometry and analyzed for GFP fluorescence (FL1 channel) after 48 h, as described in [32]. Two independent experiments were performed in triplicate.…”

Section: Assessment Of Transgene Expressionmentioning

confidence: 99%

“…Complexes were formed as for DLS measurements with either labelled or unlabelled gWIZ (control) and added to cells 30 min after vortexing (0.5 lg DNA well À1 ). After 5 h, the medium was replaced, cells were prepared for flow cytometry 24 h post-transfection and analyzed for Cy3 fluorescence (FL2 channel [32]). …”

Section: Dynamic Light Scatteringmentioning

confidence: 99%

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Specific effects of PEGylation on gene delivery efficacy of polyethylenimine: Interplay between PEG substitution and N/P ratio

Fitzsimmons

Uludağ

2012

Acta Biomaterialia

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Section: Sizes Of Pei-peg Polyplexessupporting

confidence: 90%

“…The sources of all plasmids and tissue culture reagents were described previously [32]. SYBR Green I was purchased from Invitrogen (Carlsbad, CA).…”

Section: Methodsmentioning

confidence: 99%

Section: Assessment Of Transgene Expressionmentioning

confidence: 99%

Section: Dynamic Light Scatteringmentioning

confidence: 99%

See 2 more Smart Citations

Specific effects of PEGylation on gene delivery efficacy of polyethylenimine: Interplay between PEG substitution and N/P ratio

Fitzsimmons

Uludağ

2012

Acta Biomaterialia

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“…In this study we investigated the efficiency of hydrophobe-substituted bPEI derivatives in the transfection of these cells. Different fatty acid-based substitutes have been investigated, namely stearic acid (St, C18 chain), linoleic acid (LA, C18:2 (6, 9) chain), and propionic acid (PrA, C3 chain), already reported to improve the transfection activity of bPEI on other cell types such as human embryonic kidney cells (293T) [17], rat bone marrow stromal cells (rBMSC) [25] and human breast cancer cells (MDA-231 and MCF-7) [19]. Besides, two new families of bPEI derivatives, carrying α-linoleic acid (αLA, C18:3 (3, 6, 9) chain) and acrylic acid (AcA, C3:1 (1) chain), were synthesised and studied, aiming to evaluate whether an increase in the level of unsaturation in the aliphatic chain of linoleoyl-and propionyl-based substitutes (i.e., an additional double bond, in position 3 and 1, respectively) could lead to a further enhancement in the transfection efficiency of bPEI derivatives on vascular cells.…”

Section: Resultsmentioning

confidence: 99%

Hydrophobe-substituted bPEI derivatives: boosting transfection on primary vascular cells

et al. 2017

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Gene therapy targeted to vascular cells represents a promising approach for prevention and treatment of pathological conditions such as intimal hyperplasia, in-stent and post-angioplasty restenosis. In this context, polymeric non-viral gene delivery systems are a safe alternative to viral vectors but a further improvement in efficiency and cytocompatibility is needed to improve their clinical success. Herein, a library of 24 branched polyethylenimine (bPEI) derivatives modified with hydrophobic moieties was synthesised, characterised and tested in vitro on primary vascular cells, aiming to identify delivery agents with superior transfection efficiency and low cytotoxicity. Low molecular weight PEIs (0.6, 1.2 and 2 kDa) were grafted with long (C18) and short (C3) aliphatic chains, featuring different unsaturation degrees and degrees of substitution. 0.6 kDa bPEI-based derivatives were generally ineffective in transfection on vascular smooth muscle cells (VSMCs), while among the other derivatives some promising vectors were identified. Forcing polyplexes on the cell surface by means of centrifugation invariably boosted transfection levels but increased cytotoxicity as well. Of note, a propionyl-substituted derivative (PEI2-PrA1, C3:0) was the most effective on both VSMCs and endothelial cells (ECs), with higher and more sustained gene expression in combination with markedly lower cytotoxicity with respect to the gold standard 25 kDa bPEI. In addition, a linoleoyl-substituted derivative (PEI1.2-LA6, C18:2) owing to its high efficiency in VSMCs and relative inefficacy in ECs, combined with tolerable cytotoxicity was proposed as a vector for specific VSMCs targeting.

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A Fluorophore‐Labeled Lysine Dendrimer with an Oxo‐Anion‐Binding Motif for Tracking Gene Transfection

et al. 2023

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A transfection vector based on a peptide dendrimer (1) has been developed and its abilities for DNA binding and transport have been investigated. By attaching a fluorophore to the vector system (1*), several steps in the transfection process could be monitored directly. As DLS and AFM studies showed, the labeled vector 1* condensed DNA into tightly packed aggregates able to enter eukaryotic cells. Co-localization experi-ments revealed that the ligand/plasmid complex is taken up by the endosomal pathway followed by an endosomal escape or lysosomal degradation. Afterwards, the plasmid DNA seems to enter the nucleus due to a breakdown of the nuclear envelope during mitosis, as only cells that have recently undergone mitosis showed H2B-GFP expression.

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Improved transfection efficiency of an aliphatic lipid substituted 2 kDa polyethylenimine is attributed to enhanced nuclear association and uptake in rat bone marrow stromal cell

Abstract: Improved transfection efficiency of PEI2LA was attributed to enhanced association with the nucleus, which may be a result of hydrophobic interaction between the lipid moieties on the modified lipopolymer and the nuclear membrane.

Cited by 38 publications

References 36 publications

Specific effects of PEGylation on gene delivery efficacy of polyethylenimine: Interplay between PEG substitution and N/P ratio

Specific effects of PEGylation on gene delivery efficacy of polyethylenimine: Interplay between PEG substitution and N/P ratio

Hydrophobe-substituted bPEI derivatives: boosting transfection on primary vascular cells

A Fluorophore‐Labeled Lysine Dendrimer with an Oxo‐Anion‐Binding Motif for Tracking Gene Transfection

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