Polylysine-based transfection systems utilizing receptor-mediated delivery

Zauner, Wolfgang; Ogris, Manfred; Wagner, Ernst

doi:10.1016/s0169-409x(97)00110-5

Cited by 471 publications

(292 citation statements)

References 135 publications

Supporting

Mentioning

285

Contrasting

Unclassified

Order By: Relevance

“…It is generally speculated that the uptake amounts of complexes are higher when complexes are more condensed with polycations and close to charge neutral or positively charged. 6 On the other hand, the trend in our system of reduced gene expression for the more positively charged complexes is presumably due to the possibility that the association extent of LLO-s-s-PN with PN/pDNA decreases beyond the charge neutrality point, since free PN complexes more tightly with pDNA. The binding affinity of LLO-s-s-PN to pDNA is most likely lower than that of PN to pDNA due to the bulky LLO moiety; this is supported by the data that the LLO-ss-PN elutes from the cation exchange heparin resin at 1.3 M salt concentration, whereas PN-s-s-pyridine elutes at 1.8 M (Figure 1b).…”

Section: Discussionmentioning

confidence: 78%

“…In order to increase the cellular uptake efficiency, cationic polymers and lipids, often with targeting moieties, have been typically used to neutralize and compact polyanionic, large molecular weight plasmid DNA (pDNA). [1][2][3][4][5][6] Although these approaches improve the extent of total cellular uptake of pDNA, the low transfection efficiencies and limited effectiveness of the existing non-viral gene delivery systems are largely due to the subsequent cellular processes following the uptake: the transport of genes across cell membranes into the cytosol and eventually into the nucleus. Since most existing nonviral gene delivery schemes deliver pDNA to the endosomal/lysosomal compartments, likely resulting in rapid degradation of pDNA without overcoming the membrane barrier, significant studies have been devoted to investigating new agents that can breach the membrane barrier, particularly the membranes of the endocytic compartment.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Enhanced cytosolic delivery of plasmid DNA by a sulfhydryl-activatable listeriolysin O/protamine conjugate utilizing cellular reducing potential

Saito

2003

Gene Ther

View full text Add to dashboard Cite

Listeriolysin O (LLO), a sulfhydryl-activated pore-forming protein from Listeria monocytogenes, was tested and utilized for promoting plasmid DNA (pDNA) delivery into the cytosol of cells in culture. To render pDNA-complexing capability to LLO, the unique cysteine 484 of LLO was conjugated to polycationic peptide protamine (PN) at a 1:1 molar ratio through a reversible, endosome-labile disulfide bond. The sulfhydryl-oxidized LLO construct, LLO-s-s-PN, completely lacked its pore-forming activity, yet regained its original activity upon reduction. The enhanced cytosolic delivery using this construct therefore relies on the requisite reduction of the disulfide bond in LLO-s-s-PN by endogenous cellular reducing capacity. Condensed PN/pDNA complexes incorporating LLO-s-s-PN were tested for their enhanced gene delivery capability monitoring reporter gene expression in HEK293, RAW264.7, P388D1 cell lines and bone-marrowderived macrophages in the presence of serum. Dramatic enhancement was observed for all tested complexes with varying weight ratios. The effect was most prominent at 0.64-0.80 (w/w) of PN/pDNA upon replacing 1-4% of PN with LLO-s-s-PN, resulting in approximately three orders of magnitude higher luciferase expression compared to PN/ pDNA without apparent toxicity. These results demonstrate that incorporation of endosomolytic LLO into pDNA delivery systems in a controlled fashion is a promising approach of enhancing delivery into the cytosol of target cells in gene delivery strategies.

show abstract

Section: Discussionmentioning

confidence: 78%

Section: Introductionmentioning

confidence: 99%

Enhanced cytosolic delivery of plasmid DNA by a sulfhydryl-activatable listeriolysin O/protamine conjugate utilizing cellular reducing potential

Saito

2003

Gene Ther

View full text Add to dashboard Cite

show abstract

“…High molecular weight, positively charged polyelectrolytes such as poly(lysine) are known to exhibit cytotoxicity in vitro because of interactions between the polyelectrolytes and cell membrane phospholipids resulting in disruption of the cellular membrane structure (21,22). Therefore, the cytotoxicity of each particle series was determined by using the 3-(4,5-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) cell viability assay (Fig.…”

Section: Resultsmentioning

confidence: 99%

The effect of particle design on cellular internalization pathways

Gratton

Ropp

Pohlhaus

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

2,632

2,086

View full text Add to dashboard Cite

The interaction of particles with cells is known to be strongly influenced by particle size, but little is known about the interdependent role that size, shape, and surface chemistry have on cellular internalization and intracellular trafficking. We report on the internalization of specially designed, monodisperse hydrogel particles into HeLa cells as a function of size, shape, and surface charge. We employ a top-down particle fabrication technique called PRINT that is able to generate uniform populations of organic micro-and nanoparticles with complete control of size, shape, and surface chemistry. Evidence of particle internalization was obtained by using conventional biological techniques and transmission electron microscopy. These findings suggest that HeLa cells readily internalize nonspherical particles with dimensions as large as 3 m by using several different mechanisms of endocytosis. Moreover, it was found that rod-like particles enjoy an appreciable advantage when it comes to internalization rates, reminiscent of the advantage that many rod-like bacteria have for internalization in nonphagocytic cells.PRINT ͉ shape ͉ size ͉ surface charge

show abstract

“…Alternatively, non-viral synthetic vectors, though generally nonimmunogenic, show lower transfection efficiencies and significant cytotoxicity at high or repetitive doses. Among them polycationic polymers based on polyethylene imine (PEI) [3], dendrimers [4], chitosan derivatives [5,6] and polylysine [7] form nanometric complexes (polyplexes) with pDNA and siRNA have already demonstrated to enhance the uptake of the associated nucleotides [4]. But despite their ability to enhance the cellular uptake of condensed siRNA, they still exhibit low knockdown efficiency due to insufficient siRNA release [8].…”

Section: Introductionmentioning

confidence: 99%

Cellular delivery of polynucleotides by cationic cyclodextrin polyrotaxanes

Dandekar

Jain

Keil

et al. 2012

Journal of Controlled Release

View full text Add to dashboard Cite

Cationic polyrotaxanes, obtained by temperature activated threading of cationic cyclodextrin derivatives onto water soluble cationic polymers (ionenes), form metastable nanometric polyplexes with pDNA and combinations of siRNA with pDNA.Because of their low toxicity, the polyrotaxane polyplexes constitute a very interesting system for the transfection of polynucleotides into mammalian cells. The complexation of Cy3-labeled siRNA within the polyplexes was demonstrated by fluorescence correlation spectroscopy. The uptake of the polyplexes (red) was imaged by confocal fluorescence microscopy using the A549 cell line as a model (blue: nuclei, green: membranes). The results prove the potential of polyrotaxanes for further investigations involving knocking down genes of therapeutic interest.

show abstract

Polylysine-based transfection systems utilizing receptor-mediated delivery

Cited by 471 publications

References 135 publications

Enhanced cytosolic delivery of plasmid DNA by a sulfhydryl-activatable listeriolysin O/protamine conjugate utilizing cellular reducing potential

Enhanced cytosolic delivery of plasmid DNA by a sulfhydryl-activatable listeriolysin O/protamine conjugate utilizing cellular reducing potential

The effect of particle design on cellular internalization pathways

Cellular delivery of polynucleotides by cationic cyclodextrin polyrotaxanes

Contact Info

Product

Resources

About