Flotillin-dependent endocytosis and a phagocytosis-like mechanism for cellular internalization of disulfide-based poly(amido amine)/DNA polyplexes

Vercauteren, Dries; Piest, Martin; Aa, L.J. van der; Soraj, Monerah Al; Jones, Arwyn Tomos; Engbersen, Johan F.J.; Smedt, Stefaan C. De; Braeckmans, Kevin

doi:10.1016/j.biomaterials.2010.12.045

Cited by 87 publications

(100 citation statements)

References 81 publications

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“…As naked nucleic acids are negatively charged, they are complexed with cationic polymers or cationic liposomes to increase the cellular delivery. These complexes can take different endocytic pathways to enter the cells, dependent on their size and surface characteristics (11)(12)(13)(14). When nanoparticles fail to escape the endosomal compartment, eventual delivery to (and degradation in) the lysosomes will occur.…”

Section: Introductionmentioning

confidence: 99%

Lysosomal capturing of cytoplasmic injected nanoparticles by autophagy: An additional barrier to non viral gene delivery

Remaut

Oorschot²,

Braeckmans

et al. 2014

Journal of Controlled Release

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Autophagy or 'self-eating' is a process by which defective organelles and foreign material can be cleared from the cell's cytoplasm and delivered to the lysosomes in which degradation occurs. It remains an open question, however, whether nanoparticles that did not enter the cell through endocytosis can also be captured from the cytoplasm by autophagy. We demonstrate that nanoparticles that are introduced directly in the cytoplasm of the cells by microinjection, can trigger an autophagy response. Moreover, both polystyrene beads and plasmid DNA containing poly-ethylene-imine complexes colocalize with autophagosomes and lysosomes, as was confirmed by electron microscopy.This indicates that cytoplasmic capturing of nanoparticles can occur by an autophagy response. The capturing of nanoparticles from the cytoplasm most likely limits the time frame in which efficient nucleic acid delivery can be obtained. Hence, autophagy forms an additional barrier to non-viral gene delivery, a notion that was not often taken into account before. Furthermore, these findings urges us to reconsider the idea that a single endosomal escape event is sufficient to have the long-lasting presence of nanoparticles in the cytoplasm of the cells.

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Section: Introductionmentioning

confidence: 99%

Lysosomal capturing of cytoplasmic injected nanoparticles by autophagy: An additional barrier to non viral gene delivery

Remaut

Oorschot²,

Braeckmans

et al. 2014

Journal of Controlled Release

Self Cite

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“…Additionally, the donor and acceptor fluorophores should be chosen in order 40 to avoid overlap between the spectra of emission of donor and acceptor, which would generate 41 false positives. Finally, the hybridization of two separate oligos is from a kinetic and a 1 thermodynamic point of view less favorable 123 .…”

Section: Binary Probes 26mentioning

confidence: 99%

“…Apart from the lacO/LacI system, other naturally occurring transcription factors like zinc-finger 40 nucleases, CRISPR/Cas-based methods and transcription activator-like effectors (TALEs) areused. This last type can be designed to recognize very specific DNA sequences, which makes 1 them great tools for labeling 158,159 .…”

Section: Binary Probes 26mentioning

confidence: 99%

“…Sequence-specific polynucleotide 36 labeling, on the other hand, only occurs at a predefined primary sequence, enabling control over 37 the exact location of the labels on the target polynucleotide. In this review, the current available 38 labeling strategies for DNA-based and RNA-based polynucleotides are classified based on their 39 ability to label the nucleic acids in a random or sequence-specific way, with a special focus on 40 the pros and cons of each labeling method that is being discussed. …”

mentioning

confidence: 99%

“…The method is used for the 38 same type of experiments as described for the Label-IT nucleic acid labeling kit, but their broad 39 range of fluorophores, including far-red dyes, makes it possible to extend to in vivo applications. 40 A series of poly(glutamic acid)-based peptide coatings were for example targeted to the liver, 41 spleen, spine, sternum, and femur. DNA accumulation in these tissues was confirmed by 42…”

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confidence: 99%

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Fluorescent Labeling of Plasmid DNA and mRNA: Gains and Losses of Current Labeling Strategies

2015

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7Live-cell imaging has provided the life sciences with insights into the cell biology and 8 dynamics. Fluorescent labeling of target molecules proves to be indispensable in this regard. In 9 this review, we focus on the current fluorescent labeling strategies for nucleic acids, and in 10 particular messenger RNA (mRNA) and plasmid DNA (pDNA), which are of interest to a broad 11 range of scientific fields. By giving a background of the available techniques and an evaluation 12 of the pros and cons, we try to supply scientists with all the information needed to come to an 13 informed choice of nucleic acid labeling strategy aimed at their particular needs. 14

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siRNA Transfection with Calcium Phosphate Nanoparticles Stabilized with PEGylated Chelators

Giger¹,

Castagner²,

Räikkönen³

et al. 2012

Adv Healthcare Materials

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Despite the enormous therapeutic potential of siRNAs, their delivery is still problematic due to unfavorable biodistribution profiles and poor intracellular bioavailability. Calcium phosphate co-precipitate has been used for nearly 40 years for in vitro transfection due to its non-toxic nature and simplicity of preparation. However, rapid particle growth has largely prevented the translation of this method for in vivo purposes. It has recently been shown that bisphosphonate derivatives can physically stabilize calcium phosphate nanoparticles while still allowing for efficient cell transfection with plasmid DNA. Herein, two novel PEGylated chelating agents (PEG-alendronate and PEG-inositolpentakisphosphate) with enhanced stabilizing properties are introduced, and it is demonstrated that the bisphosphonate-stabilized nanoparticles can efficiently deliver siRNA in vitro. The nanoparticles are mainly taken up by clathrin-dependent endocytosis, and acidification of the endosomal compartment is required to release the entrapped siRNA into the cytosol. Furthermore, particle uptake enhances the inhibition of the mevalonate pathway by the bisphosphonate in macrophages.

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Flotillin-dependent endocytosis and a phagocytosis-like mechanism for cellular internalization of disulfide-based poly(amido amine)/DNA polyplexes

Cited by 87 publications

References 81 publications

Lysosomal capturing of cytoplasmic injected nanoparticles by autophagy: An additional barrier to non viral gene delivery

Lysosomal capturing of cytoplasmic injected nanoparticles by autophagy: An additional barrier to non viral gene delivery

Fluorescent Labeling of Plasmid DNA and mRNA: Gains and Losses of Current Labeling Strategies

siRNA Transfection with Calcium Phosphate Nanoparticles Stabilized with PEGylated Chelators

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